КОМПОЗИЦИИ КЛЕТОК И СПОСОБЫ ИХ ПРИМЕНЕНИЯ ДЛЯ ЛЕЧЕНИЯ СЕРДЕЧНОЙ ТКАНИ

Группа изобретений относится к медицине и касается композиции для лечения сердечной ткани, содержащей TGFβ-1, ВМР4, α-тромбин, кардиотрофин и кардиогенол С; способа получения из стволовых клеток млекопитающих, дифференцированных клеток-кардиопредшественников, включающего культивирование исходных клеток в присутствии указанной композиции; способа обеспечения сердечной ткани кардиомиоцитами, который включает введение в сердечную ткань дифференцированных клеток, полученных указанным выше способом получения. Группа изобретений обеспечивает улучшенную специфичность в регулировании транскрипции генов, необходимых для индукции дифференцирования. 5 н. и 22 з.п. ф-лы, 23 ил., 1 табл., 3 пр.

ГЕНЕРАЦИЯ ФУНКЦИОНАЛЬНЫХ БЕТА-КЛЕТОК, ПОЛУЧЕННЫХ ИЗ ЧЕЛОВЕЧЕСКОЙ ПЛЮРИПОТЕНТНОЙ СТВОЛОВОЙ КЛЕТКИ, ДЕМОНСТРИРУЮЩИХ РЕАКЦИЮ В ВИДЕ ГЛЮКОЗОЗАВИСИМОГО МИТОХОНДРИАЛЬНОГО ДЫХАНИЯ И ДВУХФАЗНОЙ СЕКРЕЦИИ ИНСУЛИНА

Настоящее изобретение относится к области биотехнологии. Предложен способ доведения незрелой панкреатической бета-клетки до зрелой панкреатической бета-клетки, экспрессирующей PDX1, NKX6.1, MAFA, UCN3 и SLC2A1. Кроме того, рассмотрен способ дифференцировки плюрипотентных стволовых клеток в зрелые бета-клетки, экспрессирующие PDX1, NKX6.1, MAFA, UCN3 и SLC2A. Данное изобретение может найти дальнейшее применение в заместительной клеточной терапии для лечения сахарного диабета. 2 н. и 41 з.п. ф-лы, 10 ил., 23 табл., 7 пр.

СПОСОБ ПОЛУЧЕНИЯ МЕГАКАРИОЦИТОВ И/ИЛИ ТРОМБОЦИТОВ ИЗ ПЛЮРИПОТЕНТНЫХ СТВОЛОВЫХ КЛЕТОК

... 1. Способ получения мегакариоцитов и/или тромбоцитов, включающий культивирование гемопоэтических клеток-предшественников, полученных из плюрипотентных стволовых клеток ex vivo в присутствии соединения, представленного формулой (I), таутомера, пролекарства или фармацевтически приемлемой соли соединения или его сольвата, и дифференцировку гемопоэтических клеток-предшественников в мегакариоциты и/или тромбоциты:,где W представляет собой заместитель, представленный формулой (Ia), или карбоксигруппу:,каждый из R, R, Rи Rнезависимо представляет собой Cалкильную группу, которая может быть замещена одним или более атомами галогена или атомом водорода,n является целым числом 0, 1, 2 или 3,Rпредставляет собой Cарильную группу, которая может быть замещена одним или более заместителями, независимо представленными V, при условии что, когда n равно 2, Rне является незамещенной пиридильной группой,Rпредставляет собой Cалкильную группу, которая можетбыть замещена одним или более атомами галогена или атомом ...

AGENTS AND METHODS FOR TREATING AND PREVENTING SEBORRHEIC KERATOSIS

Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex-vivo. Also provided herein are screening assays for an agent for inducing apoptosis using cultured seborrheic keratosis cells, and methods for treating seborrheic keratosis in a subject. Methods of treating seborrheic keratosis in a subject by administering a composition that inhibits the Akt signalling pathway by topical or systemic administration of a small molecule, peptide or RNAi agent is also provided herein. The inhibitors are Akt-1 or Akt-2 inhibitors or are pan-Akt inhibitors.

Media and methods for cell culture

The present invention provides a cell passaging medium comprising at least one agent capable of detaching from a surface a cell that is culture ...

Method for determining the cardio-generative potential of mammalian cells

This document is related to a method for determining the cardio generative potential of mammalian cells which comprises the assessment of a CARdiac generation Potential Index (CARPI) as a function of the quantification of the expression of genes of said cells. It also relates to a method for quantitatively assessing the modification of this cardio-generative potential and the cardiogenic potential of a treatment aiming at cellular differentiation.

Composition for differentiation induction of adipocyte containing stem cell-derived exosome, regeneration of adipose tissue, and skin whitening or wrinkle improvement

The present invention relates to a pharmaceutical composition for adipocyte differentiation induction and/or adipose tissue regeneration comprising, as an active ingredient, an exosome extracted from a stem cell which is differentiating into an adipocyte. The exosome is excellent in expression rate of bioactive factors influencing the differentiation into an adipocyte and has the effect of differentiating a stem cell into an adipocyte. Accordingly, the present invention can be applied to differentiation inducing agents of a stem cell, injections for tissue regeneration, fillers for cosmetic purposes, preparations for tissue engineering, etc. The invention also relates to a cosmetic composition for skin whitening, wrinkle improvement and regeneration, containing an exosome extracted from a stem cell as an active ingredient. Since the exosome contains genes, proteins, growth factors etc. associated with the proliferation, differentiation, and regeneration of stem cells; is a purified component ...

CHEMICAL REPROGRAMMING OF HUMAN GLIAL CELLS INTO NEURONS WITH SMALL MOLECULE COCKTAIL

Provided are compositions, articles and methods that relate to promoting neurogenesis or neuroregeneration in mammalian nervous system. Embodiments relate to use of groups of compounds that contain Crizotinib (Cri), Flurbiprofen, Lithium Chloride (Li), Vitamin C (VC), Ceritinib (Cer) or Pirfenidone (PFD). In certain implementations glial cells are converted into functional neurons.

METHOD FOR SERUM-FREE CULTURE OF CHONDROCYTES AND SERUM-FREE CULTURE MEDIUM

... [Problem] To provide a method for serum-free culture of human cartilage cells and a serum-free culture medium. [Solution] A method for serum-free culture of cartilage cells, said method comprising: an enzymatic treatment step for treating a human cartilage cell-containing tissue with a protease; an inhibitor-treatment step for, after the enzymatic treatment step, treating the tissue with an inhibitor for the aforesaid protease; and a culture step for, after the inhibitor-treatment step, culturing the tissue in a serum-free culture medium that contains kartogenin and/or SAG, ITS, FGF2 and hydrocortisone. A serum-free culture medium for culturing cartilage cells, said serum-free culture medium containing kartogenin and/or SAG, ITS, FGF2 and hydrocortisone.

IMPROVED METHODS OF PRODUCING RPE CELLS AND COMPOSITIONS OF RPE CELLS

The present invention provides improved methods for producing RPE cells from human embryonic stem cells or from other human pluripotent stem cells. The invention also relates to human retinal pigmented epithelial cells derived from human embryonic stem cells or other human multipotent or pluripotent stem cells. hRPE cells derived from embryonic stem cells are molecularly distinct from adult and fetal-derived RPE cells, and are also distinct from embryonic stem cells. The hRPE cells described herein are useful for treating retinal degenerative diseases.

METHOD OF WASHING ADHERENT CELL USING TREHALOSE-CONTAINING CELL-WASHING SOLUTION

It is to provide a method of washing an adherent cell, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; a cell-washing solution used for the washing method; a method of producing a cell suspension for transplantation using the cell-washing solution; and a kit comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to a physiological aqueous solution to prepare a cell-washing solution containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solution can be used to wash an adherent cell before detaching the adherent cell from a culture vessel by proteolytic enzyme treatment to suppress (inhibit) cell death (apoptosis or the like) due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing (inhibiting) ...

METHODS FOR DIAGNOSING AND/OR TREATING STERILITY

The present invention concerns a polypeptide comprising the N-terminal fragment of a cystatin, and/or a cystatin expression inducer for use for the treatment of sterility and/or infertility, and a method of diagnosing and/or predicting sterility and/or infertility of a subject and/or of a couple of subjects, which method comprises measuring the level of at least one cystatin in a biological sample of a male subject and/or measuring the level of at least one cystatin in a biological sample of a female subject, and optionally measuring further the level of at least one cathepsin in a biological sample of said male of female subject.

COMPOSITION FOR MAINTAINING FUNCTION OF PLATELETS

A composition for maintaining a function of platelets, the composition comprising, as an active ingredient, a compound represented by the following general formula (I) or a salt thereof, or a solvate thereof : (see formula I) wherein X represents a phenylene group; Y represents any one of a hydrogen atom and - (CH2)mR1; wherein m represents an integer of any one of 0 to 4; and R1 is any one of -NR5COR2, -NR5SO2R2, and -NR3R4; wherein R2 represents any one of a C1 to C6 alkyl group, an aryl group, a C1 to C6 alkoxy group, and the like; R3 and R4 represent a C1 to C6 alkyl group or the like; and R5 represents any one of a hydrogen atom, a C1 to C6 alkyl group, and the like; and Z represents any one of a hydrogen atom and a C1 to C6 alkyl group.

EX VIVO MATURATION OF ISLET CELLS

The invention relates to methods for promoting maturation of islet cells from pre-weaned mammals for the purpose of optimizing the islets for their use as donor tissue for xenotransplantation. The method of the invention removes the pancreas from donor animals and reduces the pancreas tissue to fragments that are greater than the size of an intact islet while retaining islets in their whole, insulin-producing condition. The method of the invention also serially cultures the digested tissue in novel maturation media that enhance the glucose responsiveness of the cultured islets, and selects islets that are sufficiently glucose-responsive for use in transplantation procedures.

METHOD OF PRODUCING CELLS - PRECURSORS OF MATURE LIVER

METHOD OF PRODUCING CELLS OF - PRECURSORS OF MATURE LIVER

COMPOSITION FOR NY TO INCREASE THE EFFICIENCY OF ENGRAFTMENT HEMOPOIETIC STEM CELLS AFTER TRANSPLANTATION

PRODUCTION OF YOGHURT

Method for producing adult liver progenitor cells

Pancreatic islet separation method, and protective solution for protecting pancreatic islet tissue

The disclosed pancreatic islet separation method comprises: an injection step wherein a protective solution is injected into the pancreatic duct of an extracted pancreas; a preservation step wherein the pancreas is immersed into an immersion fluid and preserved; a digestion step wherein the pancreas is broken down and pancreatic tissue is obtained; and a purification step wherein the pancreatic tissue is immersed in a purification solution and pancreatic islets are obtained. The digestion step comprises: an enzyme injection step wherein an enzyme solution containing digestion enzymes is injected into the pancreas; a digestion initiation step wherein the digestion enzymes are activated; a digestion termination step wherein the digestion enzymes are de-activated; and a collection step wherein the broken down pancreatic tissue is collected. The pancreatic islet separation method is characterised in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation ...

COMPOSITION FOR GROWTH MEDIA OF MAMMALS EARLY EMBRYO COMPRISING SECRETORY LEUKOCYTE PEPTIDASE INHIBITOR AND METHOD FOR CULTURING MAMMALS EARLY EMBRYO IN VITRO TO KEEP NORMAL DEVELOPMENTAL SPEED USING THE SAME

MEASURING METHOD FOR CARDIO-GENERATIVE POTENTIAL OF MAMMALIAN CELLS

METHOD FOR PRODUCING MEGAKARYOCYTES AND/OR PLATELETS FROM PLURIPOTENT STEM CELLS

EX VIVO PRODUCTION OF PLATELETS FROM HEMATOPOIETIC STEM CELLS AND THE PRODUCT THEREOF

The present invention relates to a method of producing platelets (characterized by CD41+ and CD42+) from hematopoietic stem cells ex vivo. The invention further relates to a composition comprising the ex vivo produced platelets and a reagent comprising an aryl hydrocarbon antagonist and its use in autologous or allogeneic transfusion for the treatment of thrombocytopenia, thereby treating disease and conditions that caused the thrombocytopenia.

Method for promoting generation of stem cell-derived exosome by using thrombin

The present invention relates to a method of promoting generation of exosomes from stem cell by using thrombin. The method according to the present invention has superior effects of promoting generation of stem cell-derived exosomes and thus exosomes can be more efficiently obtained thereby compared to conventionally known methods. In addition, the method can be useful for related research.

EPITHELIAL TUMOR CELL CULTURES

Provided are cell cultures and related methods for the enrichment and conditional reprogramming of patient-derived, primary epithelial tumor cells from cancer-tissue originated spheroids (CTOSs). Also provided are methods of evaluating the responsiveness of the epithelial tumor cells to one or more therapeutic agents. 1. A cell culture medium , comprising(a) human ex vivo-derived cancer-tissue originated spheroids (CTOSs) which are dissociated into a substantially single cell suspension, wherein the CTOSs comprise human epithelial tumor cells,(b) feeder cells; and(c) a defined cell culture medium that comprises at least one Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor, optionally wherein the defined cell culture medium does not comprise or is substantially free of a Bone Morphogenetic Protein (BMP) inhibitor such as Noggin and a Wnt/β-catenin signaling agonist such as R-spondin-1; and optionally(d) supplemental serum albumin (SA),wherein the cell culture medium provides at least about 10% proliferation of the epithelial tumor cells within about 14 days following co-culture of the dissociated CTOSs with (b) and (c).2. The cell culture medium of claim 1 , wherein the human ex vivo-derived CTOSs are cultured from a tumor-containing sample removed from a human patient with cancer claim 1 , optionally selected from a surgical sample claim 1 , a biopsy sample claim 1 , a pleural effusion sample claim 1 , and an ascetic fluid sample.3. The cell culture medium of or claim 1 , wherein the human patient has a cancer selected from colon cancer claim 1 , lung cancer claim 1 , gastric cancer claim 1 , and breast cancer.4. The cell culture medium of or claim 1 , wherein the epithelial tumor cells substantially retain the clonal diversity of the tumor-containing sample removed from the human patient within about 14 days following co-culture of the dissociated CTOSs with (b) and (c).5. The cell culture medium of any one of - claim 1 , wherein the human ...

METHOD FOR DIFFERENTIATION OF OCULAR CELLS AND USE THEREOF

Provided herein are methods of producing a photoreceptor precursor (PRP) cell population derived from stem cells. Further provided herein are methods of using the PRP cell populations, such as for therapeutics.

КОМПОЗИЦИИ КЛЕТОК И СПОСОБЫ ИХ ПРИМЕНЕНИЯ ДЛЯ ЛЕЧЕНИЯ СЕРДЕЧНОЙ ТКАНИ

... 1. Композиция, содержащая TGFβ-1, BMP4, α-тромбин, соединение, выбранное из группы, состоящей из кардиотрофина и IL-6, и соединение, выбранное из группы, состоящей из кардиогенола C и ретиноевой кислоты. ! 2. Композиция по п.1, которая содержит TGFβ-1, BMP4, α-тромбин, кардиотрофин и кардиогенол C. ! 3. Композиция по п.1, которая содержит, по меньшей мере, одно соединение, выбранное из группы, состоящей из FGF-2, IGF-1, активина-A, TNF-α, FGF-4, LIF, VEGF-А и их комбинаций. ! 4. Композиция по любому из предшествующих пунктов, которая содержит FGF-2, IGF-1 и активин-A. ! 5. Композиция по п.1, которая содержит активин-A, FGF-2, IL-6, IGF-1 и ретиноевую кислоту. ! 6. Композиция по п.1, которая не содержит, по меньшей мере, одного соединения, выбранного из группы, состоящей из TNF-α, FGF-4, LIF и VEGF-A. ! 7. Композиция по п.1, в которой когда одно соединение присутствует в указанной композиции, оно присутствует в количестве между 1 и 5 нг TGFβ-1 на мл, между 1 и 10 нг BMP4 на мл, между 0,5 ...

ВЫСОКОБЕЗОПАСНЫЙ СПОСОБ ПОЛУЧЕНИЯ ОЧИЩЕННЫХ ФРАКЦИЙ СТВОЛОВЫХ КЛЕТОК

... 1. Способ получения фракций стволовых клеток, имеющих происхождение из липидной ткани, включающий стадии:(a) сбора или получения образца липидной ткани, содержащей стволовые клетки;(b) промывания образца со стадии (a) подходящим водным буфером;(c) инкубирования образца со стадии (b) с ферментом, способным перерабатывать липидную ткань и извлекать из нее стволовые клетки;(d) извлечения водной фазы из продукта со стадии (c);(e) очистки водной фазы, полученной на стадии (d);(f) титрования водной фазы, полученной на стадии (e), и, если необходимо, ее разбавление, чтобы получить конечную фракцию стволовых клеток с желаемой концентрацией и объемом,в котором материал, содержащий стволовые клетки, обрабатывают в шприце на всем протяжении, по меньшей мере, стадий: (a), (b) и (c), указанный шприц, выполняет функции собирающего устройства, фазового разделителя и технологического реактора.2. Способ по п.1, в котором шприц имеет объем заполнения, составляющий от 20 до 100 мл.3. Способ по п.1, в котором ...

Verfahren zur Bildung von Nierentubuli

Die Erfindung betrifft ein Verfahren zur Bildung von Nieren-Tubuli durch Einbettung von einzelnen Nieren-Zellen in ein synthetisches Hydrogel, basierend auf Polyethylenglykol als Komponente, und die Kultivierung der Zellen bis zur Bildung von Tubulistrukturen. Die Kultivierung kann solange fortgesetzt werden, bis die erhaltenen Tubulistrukturen in Größe, Struktur und Morphologie und Funktionalität adulten humanen Nierentubuli entsprechen oder zumindest ähneln.

Medium for direct differentiation of pluripotent stem cell-derived mesenchymal stem cell, method for preparing mesenchymal stem cell by using same, and mesenchymal stem cell prepared thereby

The present invention relates to a medium for direct differentiation of embryonic stem cell-derived mesenchymal stem cells, a method for preparing mesenchymal stem cells by using same, mesenchymal stem cells prepared thereby, and a cell therapy product comprising the same mesenchymal stem cells. In a medium composition and a method according to an embodiment, mesenchymal stem cells can be prepared at high yield within a short period of time. In addition, the method is simple in preparation procedure because of the absence of an embryoid body formation step and allows homogeneous cells to be prepared, thus advantageously providing a cell therapy product within a reduced period of time, compared to conventional methods.

Method for determining the cardio-generative potential of mammalian cells

This document is related to a method for determining the cardio generative potential of mammalian cells which comprises the assessment of a CARdiac generation Potential Index (CARPI) as a function of the quantification of the expression of genes of said cells. It also relates to a method for quantitatively assessing the modification of this cardio-generative potential and the cardiogenic potential of a treatment aiming at cellular differentiation.

Chemical reprogramming of human glial cells into neurons with small molecule cocktail

Provided are compositions, articles and methods that relate to promoting neurogenesis or neuroregeneration in mammalian nervous system. Embodiments relate to use of groups of compounds that contain Crizotinib (Cri), Flurbiprofen, Lithium Chloride (Li), Vitamin C (VC), Ceritinib (Cer) or Pirfenidone (PFD). In certain implementations glial cells are converted into functional neurons.

Method for culturing pluripotent stem cells

Provided is a method whereby de-undifferentiated cells that occur in a colony during culturing pluripotent stem cells can be removed. One embodiment of the present invention relates to a method for culturing pluripotent stem cells, said method comprising conducting the cell culture in the presence of a substance capable of inhibiting intracellular adhesion. Another embodiment thereof relates to a method for removing de-undifferentiated cells that may possibly occur or have occurred in a colony during culturing pluripotent stem cells, said method comprising conducting the cell culture in the presence of a substance capable of inhibiting intracellular adhesion. Another embodiment thereof relates to a method for maintaining an undifferentiated state of pluripotent stem cells, said method comprising conducting the cell culture in the presence of a substance capable of inhibiting intracellular adhesion.

COMPOSITION FOR USE IN INCREASING ENGRAFTMENT EFFICACY OF HAEMATOPOETIC STEM CELLS AFTER TRANSPLANTATION

The present invention provides the new use of composition comprising at least one inhibitor of dipeptidyl peptidase IV (DPP-IV) for increasing migration and homing of haematopoetic progenitor cells in stem cell transplanted recipients, wherein said haematopoetic stem and/or progenitor cells had been treated in vitro with an engraftment enhancing compound, specifically with a prostacyclin analogue and a cAMP enhancer before transplantation.

COMPOSITIONS AND METHODS FOR USING CELLS TO TREAT HEART TISSUE

This document relates to compositions containing cardiogenic factors, to methods to obtain cells by culturing initial cells in the presence of such factors; and methods of administering the obtained cells to heart tissue.

METHOD FOR CULTURING PLURIPOTENT STEM CELLS

Provided is a method whereby de-undifferentiated cells that occur in a colony during culturing pluripotent stem cells can be removed. One embodiment of the present invention relates to a method for culturing pluripotent stem cells, said method comprising conducting the cell culture in the presence of a substance capable of inhibiting intracellular adhesion. Another embodiment thereof relates to a method for removing de-undifferentiated cells that may possibly occur or have occurred in a colony during culturing pluripotent stem cells, said method comprising conducting the cell culture in the presence of a substance capable of inhibiting intracellular adhesion. Another embodiment thereof relates to a method for maintaining an undifferentiated state of pluripotent stem cells, said method comprising conducting the cell culture in the presence of a substance capable of inhibiting intracellular adhesion.

METHODS FOR PRODUCTION OF PLATELETS FROM PLURIPOTENT STEM CELLS AND COMPOSITIONS THEREOF

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.

METHOD OF WASHING ADHERENT CELL USING TREHALOSE-CONTAINING CELL-WASHING SOLUTION

It is to provide a method of washing an adherent cell, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; a cell-washing solution used for the washing method; a method of producing a cell suspension for transplantation using the cell-washing solution; and a kit comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to a physiological aqueous solution to prepare a cell-washing solution containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solution can be used to wash an adherent cell before detaching the adherent cell from a culture vessel by proteolytic enzyme treatment to suppress (inhibit) cell death (apoptosis or the like) due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing (inhibiting) ...

HIGH-SAFETY PROCESS FOR THE PREPARATION OF PURIFIED STEM CELL FRACTIONS

A highly safe procedure for the preparation of purified stem cell fractions of lipid origin is herein described, in which the use of a specially designed single collecting device, reduces the number of passages and manipulations undergone by stem cell-containing material, reducing to a minimum the risks of contamination, material loss, and inadvertent exchange of samples, and further simplifying the interface and cooperation between personnel recovering the raw material and those expert in stem cell isolation.

심장 조직을 처치하는 세포를 이용하는 조성물 및 방법

... 본 문헌은 심장 인자를 함유하는 조성물, 상기 인자의 존재시 초기 세포를 배양함으로써 세포를 얻는 방법, 및 상기 방법에 의해 얻은 세포에 관한 것이다.

PANCREATIC ISLET SEPARATION METHOD, AND PROTECTIVE SOLUTION FOR PROTECTING PANCREATIC ISLET TISSUE

The disclosed islet isolation method comprises: an injection step of injecting a preservation solution into the pancreatic duct of an excised pancreas; a preservation step of immersing the pancreas into an immersion fluid for preservation; a digestion step of breaking down the pancreas to provide pancreatic tissue; and a purification step of immersing the pancreatic tissue in a purification solution to provide islets. The digestion step consists of: an enzyme injection step of injecting an enzyme solution containing a digestion enzyme into the pancreas; a digestion initiation step of activating the digestion enzyme; a digestion termination step of inactivating the digestion enzyme; and a collection step of collecting the broken-down pancreatic tissue. The islet isolation method is characterized in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation step, the neutrophil elastase inhibitor is present inside the pancreas at the time point of starting ...

PRODUCTION METHODS FOR MEGAKARYOCYTES AND PLATELETS

An object of the present invention is to provide a method of efficiently producing a maturated megakaryocytic cell line from hematopoietic progenitor cells. The present invention provides a method for producing megakaryocytes from hematopoietic progenitor cells, comprising (i) forcibly expressing an apoptosis suppression gene and an oncogene in hematopoietic progenitor cells and culturing the cells, and (ii) arresting forced expression of the apoptosis suppression gene and the oncogene and culturing the hematopoietic progenitor cells.

Cosmetic Composition Containing Exosomes Extracted from Stem Cell for Skin Whitening, Antiwrinkle or Regeneration

A cosmetic composition for skin whitening, wrinkle improvement or skin regeneration includes, as an active ingredient, exosomes derived from stem cells comprising proliferating stem cells.

NOVEL METHOD FOR OBTAINING T CELLS FROM PLURIPOTENT STEM CELLS, AND USES THEREOF

A method for obtaining a population of T cells from pluripotent stem cells, which includes a first step of obtaining hematopoietic stem cells and a second step of obtaining T cells. Also, the cell populations thus obtained according to this method and these cell populations for use as a medicament.

PHARMACEUTICAL COMPOSITION FOR TREATING CEREBROVASCULAR DISEASES, CONTAINING STEM CELL-DERIVED EXOSOME AS ACTIVE INGREDIENT

The present invention relates to a pharmaceutical composition for treating cerebrovascular diseases including stem cell-derived exosomes as an active ingredient. The stem cell-derived exosomes according to the present invention have superior nerve cell protective effects, such as inhibition of cerebral ventricular distention, reduction of hydrocephalus, and inhibition of nerve cell death and cellular inflammation in an intraventricular hemorrhage (IVH) animal model, and thus, can be useful in treating cerebrovascular diseases including intraventricular hemorrhage, etc.

HIGH-SAFETY PROCESS FOR THE PREPARATION OF PURIFIED STEM CELL FRACTIONS

A highly safe procedure for the preparation of purified stem cell fractions of lipid origin is herein described, in which the use of a specially designed single collecting device, reduces the number of passages and manipulations undergone by stem cell-containing material, reducing to a minimum the risks of contamination, material loss, and inadvertent exchange of samples, and further simplifying the interface and cooperation between personnel recovering the raw material and those expert in stem cell isolation. 2. Process according to claim 1 , wherein the syringe has a filling volume comprised between 20 and 100 mL.3. Process according to claim 1 , wherein the syringe is provided with one or more marks to indicate the optimal filling volume(s) claim 1 , and/or with areas for writing or labelling.4. Process according to claim 1 , wherein the lipid material is a lipoaspirate.5. Process according to claim 1 , wherein the step (a) and (b-d) respectively claim 1 , are performed by two different operators at locations remote from each other.6. Process according to claim 1 , wherein in step (b) the buffer is a PBS buffer supplemented with enzyme nutrients.7. Process according to claim 1 , in which the steps (b) and/or (c) include orienting the syringe downwards (needle down) or upwards (needle up) claim 1 , followed by ejecting the phase proximal to the needle.8. Process according to claim 1 , wherein in step (c) the enzyme is a liberase claim 1 , and the incubation is performed for 20 to 80 minutes claim 1 , at a temperature from 30 to 45° C.9. Process according to claim 1 , wherein in step (d) claim 1 , the incubated mixture is mixed with an albumin-containing solution claim 1 , then the lipid phase is discarded and the aqueous phase is recovered for the subsequent process steps.10. Process according to claim 1 , where the stem cells-containing material is further maintained within the said syringe during one or more of the steps (d)-(e).11. Process according to claim 1 ...

ВЫСОКОБЕЗОПАСНЫЙ СПОСОБ ПОЛУЧЕНИЯ ОЧИЩЕННЫХ ФРАКЦИЙ СТВОЛОВЫХ КЛЕТОК

Изобретение относится к биохимии. Описан способ получения фракций стволовых клеток, имеющих происхождение из липидной ткани, включающий стадии: (a) сбора или получения образца липидной ткани, содержащей стволовые клетки; (b) промывания образца со стадии (а) подходящим водным буфером; (c) инкубирования образца со стадии (b) с ферментом, способным перерабатывать липидную ткань и извлекать из нее стволовые клетки; (d) инактивирования фермента, использованного на стадии (с), и извлечения водной фазы из продукта со стадии (с); (e) очистки водной фазы, полученной на стадии (d); (f) титрования водной фазы, полученной на стадии (е), и, если необходимо, ее разбавления, чтобы получить конечную фракцию стволовых клеток с желаемой концентрацией и объемом, в котором материал, содержащий стволовые клетки, обрабатывают в шприце на всем протяжении по меньшей мере стадий: (а), (b) и (с), указанный шприц, выполняет функции собирающего устройства, фазового разделителя и технологического реактора. Изобретение ...

КОМПОЗИЦИЯ, ВКЛЮЧАЮЩАЯ ЭКЗОСОМУ, ПОЛУЧЕННУЮ ИЗ СТВОЛОВЫХ КЛЕТОК, ДЛЯ ИНДУКЦИИ АДИПОГЕННОЙ ДИФФЕРЕНЦИРОВКИ, РЕГЕНЕРАЦИИ ЖИРОВОЙ ТКАНИ, ОТБЕЛИВАНИЯ КОЖИ ИЛИ КОРРЕКЦИИ МОРЩИН

Группа изобретений относится к биотехнологии, а именно к композиции для индукции дифференцировки стволовых клеток в адипоциты. Композиция для индукции дифференцировки стволовых клеток в адипоциты для регенерации жировых тканей содержит в качестве ингредиента экзосомы, полученные из стволовых клеток, полученных из жировой ткани и дифференцирующихся в адипоциты, где экзосомы характеризуются тем, что содержат макрофагальный колониестимулирующий фактор (MCSF), фактор некроза опухоли-α (TNF-α), лептин, инсулин, ангиопоэтин-1 (ANGPT1) и адипонектин с молекулярной массой 30 кДа (Acrp30). Группа изобретений относится также к инъекционной форме препарата, содержащей указанную композицию. Использование данной группы изобретений позволяет применять данную композицию в качестве агентов, индуцирующих дифференцировку стволовых клеток, инъекционных форм препаратов для регенерации тканей путем экспрессии экзосомами биоактивных факторов, оказывающих воздействие на дифференцировку в адипоциты. 2 н. и 4 з.п ...

Amyloid peptide inactivating enzyme to treat alzheimer's disease

A polypeptide comprising N-terminal fragment of a cystatin for use in diagnosing and/or treating sterility

The present invention concerns a polypeptide comprising the N-terminal fragment of a cystatin, and/or a cystatin expression inducer for use for the treatment of sterility and/or infertility, and a method of diagnosing and/or predicting sterility and/or infertility of a subject and/or of a couple of subjects, which method comprises measuring the level of at least one cystatin in a biological sample of a male subject and/or measuring the level of at least one cystatin in a biological sample of a female subject, and optionally measuring further the level of at least one cathepsin in a biological sample of said male of female subject.

Agents and methods for treating and preventing seborrheic keratosis

Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex vivo. Also provided herein are screening assays using cultured seborrheic keratosis cells and methods for treating seborrheic keratosis in a subject.

Bioink composition for dermis regeneration sheet, method for manufacturing customized dermis regeneration sheet using same, and customized dermis regeneration sheet manufactured using manufacturing method

The present invention relates to a bioink composition for a dermis regeneration sheet and a method for manufacturing a customized dermis regeneration sheet using same, the bioink composition comprising: a first liquid comprising an adipose tissue-derived stromal vascular fraction, an extracellular matrix, and fibrinogen; and a second liquid comprising thrombin.

A METHOD FOR PREPARING A GROWTH FACTORS CONTAINING PLATELET RELEASATE

The present invention relates to a method for preparing a growth-factors containing platelet releasate from a fluid mammalian platelet concentrate, comprising the consecutive steps of subjecting the platelet concentrate to a pathogen reduction step to disrupt non-enveloped viruses; subjecting the platelet concentrate to an activation step to cause the platelets to release growth factors; recovering a fibrinogen depleted fluid platelet releasate; subjecting the fibrinogen depleted fluid platelet releasate to a second pathogen concentration reduction step to disrupt enveloped viruses; subjecting the platelet releasate to sterile filtering and recovering a filtrate liquid containing the growth factors. The platelet releasate obtained with the method of the present invention may be used as a therapeutic agent to enhance the proliferation of multi lineage cells in regenerative medicine and in the management of non healing wounds and resistant ulcers. The second indication is as a substitute ...

EXPRESSION VECTORS FOR CHIMERIC ENGULFMENT RECEPTORS, GENETICALLY MODIFIED HOST CELLS, AND USES THEREOF

The present disclosure relates to tandem expression cassettes encoding chimeric engulfment receptor molecules and chimeric antigen receptors/or T cell receptor binding proteins, host cells modified to include the tandem expression cassettes, and methods of making and using such receptor molecules and modified cells.

METHOD FOR PRODUCING ADULT LIVER PROGENITOR CELLS

Novel adult liver progenitor cells (called H2Stem Cells) have been have been characterized on the basis of a series of biological activities and markers. Methods for producing H2Stem Cells allow providing such cells in the form of adherent cells and three-dimensional cell clusters in suspension that can be differentiated into cells having strong liver-specific activities and/or that can be used for treating liver diseases or for evaluating the efficacy, the metabolism, and/or toxicity of a compound.

TGF.BETA. SIGNALING INDEPENDENT NAIVE INDUCED PLURIPOTENT STEM CELLS, METHODS OF MAKING AND USE

Provided is a cocktail of factors for converting/reprogramming non-naïve pluripotent stem cells into TGFß signaling-independent (TSI) naïve induced pluripotent stem cells (iPSCs). Also provided are methods for reprograming a non-naïve PSC into a TSI naïve iPSC by contacting the cell to be reprogrammed with effective amounts of compounds for a sufficient period of time to reprogram the cell into a TSI naïve iPSC.

COMPOSITION FOR MAINTAINING FUNCTION OF PLATELETS

Provided is a composition for maintaining platelet function, having as an active ingredient a pyridine derivative represented by the following general formula (I), or a salt thereof (in the formula, ring A, R1, R2, R3, and R4 represent the definitions given in the description).

METHOD FOR DETERMINING THE CARDIO-GENERATIVE POTENTIAL OF MAMMALIAN CELLS

This document is related to a method for determining the cardio generative potential of mammalian cells which comprises the assessment of a CARdiac generation Potential Index (CARPI) as a function of the quantification of the expression of genes of said cells. It also relates to a method for quantitatively assessing the modification of this cardio-generative potential and the cardiogenic potential of a treatment aiming at cellular differentiation.

PANCREATIC ISLET SEPARATION METHOD, AND PROTECTIVE SOLUTION FOR PROTECTING PANCREATIC ISLET TISSUE

The disclosed pancreatic islet separation method comprises: an injection step wherein a protective solution is injected into the pancreatic duct of an extracted pancreas; a preservation step wherein the pancreas is immersed into an immersion fluid and preserved; a digestion step wherein the pancreas is broken down and pancreatic tissue is obtained; and a purification step wherein the pancreatic tissue is immersed in a purification solution and pancreatic islets are obtained. The digestion step comprises: an enzyme injection step wherein an enzyme solution containing digestion enzymes is injected into the pancreas; a digestion initiation step wherein the digestion enzymes are activated; a digestion termination step wherein the digestion enzymes are de-activated; and a collection step wherein the broken down pancreatic tissue is collected. The pancreatic islet separation method is characterised in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation ...

PANCREATIC ISLET SEPARATION METHOD, AND PROTECTIVE SOLUTION FOR PROTECTING PANCREATIC ISLET TISSUE

Compositions and methods for producing and administering brown adipocytes

The present disclosure provides compositions comprising a hydrogel and a cell adhesion ligand that enhances the differentiation of mesenchymal stem cells into brown adipocytes, and/or enhances the maintenance of brown adipocytes. In some cases, a subject composition includes cells that are embedded within the hydrogel. The present disclosure also provides methods of making and using the subject compositions.

Method for cell culture

A method for stem or progenitor cell culture. More precisely, the invention relates to a method for cell culture using one or more IαI (inter-alpha trypsin inhibitor or inter-alpha inhibitor) protein(s) or part(s) thereof as a component in a cell culture media or a coating on a cell culture surface material. Furthermore the invention relates to a cell culture media and a cell culture coating/matrix provided with one or more IαI proteins(s) or part(s) thereof.

TWO STAGE CELLULARIZATION STRATEGY FOR THE FABRICATION OF BIOARTIFICIAL HEARTS

In some embodiments, the present disclosure pertains to a method of fabricating an artificial heart muscle (AHM) patch. In some embodiments, the method comprises obtaining and/or isolating cells from a subject. In some embodiments, the cells are primary cardiac cells. In some embodiments, the method further comprises forming a scaffold. In some embodiments, the method comprises seeding the cells in the fibrin gel scaffold. In some embodiments, the method comprises culturing the cells seeded in the fibrin gel scaffold under conditions appropriate for the formation of an artificial heart muscle (AHM) patch. In some embodiments, the present disclosure pertains to a method of fabricating a bioartificial heart (BAH). In some embodiments, the present disclosure pertains to a method of treatment of cardiac tissue injury in a subject in need thereof. In some embodiments, the method includes implanting the aforementioned artificial heart muscle patch in the injured area of the subject. In some embodiments, the present disclosure relates to a method of treating end stage cardiac disease in a subject in need thereof. 1. A method of fabricating an artificial heart muscle (AHM) patch comprising:obtaining and/or isolating cells from a subject;preparing a scaffold;seeding the cells in the prepared scaffold; andculturing the cells within the scaffold.2. The method of claim 1 , wherein the cells are primary cardiac cells.3. The method of claim 1 , wherein the cells obtained and/or isolated are neonatal cardiomyocytes.4. The method of claim 1 , wherein the subject is a mammal.5. The method of claim 1 , wherein the scaffold is formed from fibrin gel.6. The method of claim 5 , wherein the scaffold is formed by layering fibrin gel on a surface coated with a silicone elastomer.7. The method of claim 6 , wherein the silicone elastomer is polydimethylsiloxane (PDMS).8. The method of claim 5 , wherein the fibrin gel is mixed with saline and culture media containing thrombin.9. The method of ...

СПОСОБ ПОЛУЧЕНИЯ РЕЛИЗАТА ТРОМБОЦИТОВ, СОДЕРЖАЩЕГО ФАКТОРЫ РОСТА

СПОСОБ ОПРЕДЕЛЕНИЯ СЕРДЕЧНО-СОСУДИСТОГО ПОТЕНЦИАЛА В КЛЕТКАХ МЛЕКОПИТАЮЩИХ

... 1. Способ определения кардио-генеративного потенциала клеток млекопитающих, который включает оценку коэффициента кардиального генеративного потенциала (CARPI) как функции относительных количественных показателей экспрессии двух или более генов указанных клеток, причем по меньшей мере один из указанных генов выбран из группы, состоящей из генов Nkx2.5, Tbx5, MEF2C, GATA4, GATA6, Mesp1, FOG1, FOG2, Flk1, их гомологов у млекопитающих и их комбинаций.2. Способ по п.1, в котором указанную экспрессию генов измеряют на уровне матричной РНК (мРНК), функциональных РНК, таких как микроРНК, или их комбинаций.3. Способ по п.2, в котором измеряют уровень экспрессии мРНК по меньшей мере одного гена, выбранного из группы, состоящей из Nkx2.5, Tbx5, MEF2C, GATA4, GATA6, Mesp1, FOG1, FOG2, Flk1, их гомологов у млекопитающих и любых их комбинаций.4. Способ по любому из пп.1-3, в котором указанные клетки выбраны из группы, включающей соматические, половые клетки, клетки пуповинной крови, кардиальные предшественники ...

Pancreatic islet separation method, and protective solution for protecting pancreatic islet tissue

The disclosed pancreatic islet separation method comprises: an injection step wherein a protective solution is injected into the pancreatic duct of an extracted pancreas; a preservation step wherein the pancreas is immersed into an immersion fluid and preserved; a digestion step wherein the pancreas is broken down and pancreatic tissue is obtained; and a purification step wherein the pancreatic tissue is immersed in a purification solution and pancreatic islets are obtained. The digestion step comprises: an enzyme injection step wherein an enzyme solution containing digestion enzymes is injected into the pancreas; a digestion initiation step wherein the digestion enzymes are activated; a digestion termination step wherein the digestion enzymes are de-activated; and a collection step wherein the broken down pancreatic tissue is collected. The pancreatic islet separation method is characterised in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation ...

Composition for use in increasing engraftment efficacy of haematopoetic stem cells after transplantation

The present invention provides the new use of composition comprising at least one inhibitor of dipeptidyl peptidase IV (DPP-IV) for increasing migration and homing of haematopoetic progenitor cells in stem cell transplanted recipients, wherein said haematopoetic stem and/or progenitor cells had been treated in vitro with an engraftment enhancing compound, specifically with a prostacyclin analogue and a cAMP enhancer before transplantation.

COMPOSITIONS AND METHODS FOR ADOPTIVE CELL THERAPY FOR CANCER

Provided herein are compositions and methods for adoptive cell therapy comprising engineered immune cells that express a tumor antigen-targeted chimeric antigen receptor and a prodrug converting enzyme.

METHOD FOR SERUM-FREE CULTURE OF CHONDROCYTES AND SERUM-FREE CULTURE MEDIUM

... [Problem] To provide a method for serum-free culture of human cartilage cells and a serum-free culture medium. [Solution] A method for serum-free culture of cartilage cells, said method comprising: an enzymatic treatment step for treating a human cartilage cell-containing tissue with a protease; an inhibitor-treatment step for, after the enzymatic treatment step, treating the tissue with an inhibitor for the aforesaid protease; and a culture step for, after the inhibitor-treatment step, culturing the tissue in a serum-free culture medium that contains kartogenin and/or SAG, ITS, FGF2 and hydrocortisone. A serum-free culture medium for culturing cartilage cells, said serum-free culture medium containing kartogenin and/or SAG, ITS, FGF2 and hydrocortisone.

HUMAN AIRWAY STEM CELLS IN LUNG EPITHELIAL ENGINEERING

Methods of using human airway stem cells in lung epithelial engineering, optionally wherein the cells are contacted with a gamma secretase inhibitor, bioartificial airway organs produced thereby, and the use thereof, e.g., for transplantation. Also methods of treating a bio-artificial matrix with Tenascin-C and/or fibrillin 2.

IMPROVED METHODS OF PRODUCING RPE CELLS AND COMPOSITIONS OF RPE CELLS

ENDOCRINE PRECURSOR CELLS, PANCREATIC HORMONE-EXPRESSING CELLS AND METHODS OF PRODUCTION

PANCREATIC ISLET SEPARATION METHOD, AND PROTECTIVE SOLUTION FOR PROTECTING PANCREATIC ISLET TISSUE

The disclosed pancreatic islet separation method comprises: an injection step wherein a protective solution is injected into the pancreatic duct of an extracted pancreas; a preservation step wherein the pancreas is immersed into an immersion fluid and preserved; a digestion step wherein the pancreas is broken down and pancreatic tissue is obtained; and a purification step wherein the pancreatic tissue is immersed in a purification solution and pancreatic islets are obtained. The digestion step comprises: an enzyme injection step wherein an enzyme solution containing digestion enzymes is injected into the pancreas; a digestion initiation step wherein the digestion enzymes are activated; a digestion termination step wherein the digestion enzymes are de-activated; and a collection step wherein the broken down pancreatic tissue is collected. The pancreatic islet separation method is characterised in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation ...

CELL SUSPENSION AND USE THEREOF

The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration.

Compositions and methods for using cells to treat heart tissue

This document relates to compositions containing cardiogenic factors, to methods to obtain cells by culturing initial cells in the presence of such factors; and methods of administering the obtained cells to heart tissue.

METHOD FOR PRODUCING ADULT LIVER PROGENITOR CELLS

CULTURE MEDIUM COMPOSITION FOR EARLY MAMMALIAN EMBRYO GROWTH INCLUDING SECRETORY LEUKOCYTE PEPTIDASE INHIBITOR AND EARLY EMBRYO IN VITRO CULTURE METHOD USING SAME FOR MAINTAINING NORMAL GENERATION SPEED

The present invention relates to a culture medium composition for mammalian early embryo growth including a secretory leukocyte peptidase inhibitor and an early embryo vitro culture method using the same for maintaining a normal generation speed and, more specifically, to a culture medium composition and a vitro culture method using the same, capable of promoting a generation speed to a similar speed to vivo culture and improving an implantation rate in comparison to existing embryos in vitro culture. According to the present invention, the secretory leukocyte peptidase inhibitor included in the culture medium composition is able to enable the generation of an embryo in vitro culture to progress similarly to in vivo culture and to increase the expression level of various generators, thereby improving the implantation rate and early embryo generation rate in comparison to embryos cultured through a conventional in vivo culture method. The present invention is able to be applied to clinics ...

GENETICALLY MODIFIED MESENCHYMAL STEM CELLS EXPRESSING ALPHA-1 ANTITRYPSIN (AAT)

Compositions and methods for obtaining cells to treat heart tissue

This document relates to compositions containing cardiogenic factors, to methods to obtain cells by culturing initial cells in the presence of such factors; and methods of administering the obtained cells to heart tissue.

Agents and methods for treating and preventing seborrheic keratosis

Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex vivo. Also provided herein are screening assays using cultured seborrheic keratosis cells and methods for treating seborrheic keratosis in a subject.

Composition for maintaining function of platelets

A composition for maintaining a function of platelets, the composition comprising, as an active ingredient, a compound represented by the following general formula (I) or a salt thereof, or a solvate thereof: wherein X represents a phenylene group; Y represents any one of a hydrogen atom and (CH2)mR1; wherein m represents an integer of any one of 0 to 4; and R1 is any one of NR5COR2, NR5SO2R2, and NR3R4; wherein R2 represents any one of a C1 to C6 alkyl group, an aryl group, a C1 to C6 alkoxy group, and the like; R3 and R4 represent a C1 to C6 alkyl group or the like; and R5 represents any one of a hydrogen atom, a C1 to C6 alkyl group, and the like; and Z represents any one of a hydrogen atom and a C1 to C6 alkyl group.

METHODS FOR PRODUCTION OF PLATELETS FROM PLURIPOTENT STEM CELLS AND COMPOSITIONS THEREOF

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells. 1. A pharmaceutical preparation that is suitable for use in a human patient comprising at least 10platelets , wherein the preparation is substantially free of leukocytes and wherein substantially all of the platelets are functional.2. The pharmaceutical preparation of that comprises 10-10platelets claim 1 , optionally 10 claim 1 , 10 claim 1 , 10 claim 1 , 10 claim 1 , 10 claim 1 , or 10platelets.3. The pharmaceutical preparation of claim 1 , wherein the platelets have one or more of the following attributes: a mean platelet volume range of 9.7-12.8 fL; a unimodal distribution of size in the preparation; and/or a lognormal platelet volume distribution wherein one standard deviation is less than 2 μm claim 1 , (less than 1.5 μm claim 1 , less than 1 μmor less than 0.5 μm.4. The pharmaceutical preparation of claim 1 , wherein the platelets are positive for at least one of the following markers: CD41a and CD42b.5. The pharmaceutical preparation of claim 1 , wherein the platelets are human platelets.6. The pharmaceutical preparation of claim 1 , wherein at least 50% claim 1 , 60% claim 1 , 70% claim 1 , 80% or 90% of the platelets are functional for at least 2 claim 1 , 3 or 4 days after storage at room temperature.745-. (canceled)46. A method for producing a pharmaceutical preparation comprising:differentiating megakaryocytes positive for CD41a and CD42b expression in a ...

СПОСОБ ОПРЕДЕЛЕНИЯ КАРДИО-ГЕНЕРАТИВНОГО ПОТЕНЦИАЛА КЛЕТОК МЛЕКОПИТАЮЩИХ

FIELD: biotechnology. SUBSTANCE: method for cardio-generative potential determination in mammal cells is proposed, including collection of the measured levels of Nkx2.5, Tbx5, MEF2C, GATA4, GATA6, Mesp1, FOG1 genes expression in mammal cells, and cardio-generative potential coefficient (CARPI) determination as the average value of the measured levels expression of the genes of the said cells. EFFECT: good predictability of heart recovery success. 13 cl, 1 dwg, 1 tbl, 3 ex С 2 2624498 ко РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) (2 эз ааа даз0Э (51) МПК СОМ 33/50 (2006.01) С120 168 (2006.01) СОбЕ 19/24 (2011.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (21)(22) Заявка: 2011143063, 20.05.2010 (24) Дата начала отсчета срока действия патента: 20.05.2010 Дата регистрации: 04.07.2017 Приоритет(ы): (30) Конвенционный приоритет: 20.05.2009 9 РСТДЛ$2009/044751 (43) Дата публикации заявки: 27.06.2013 Бюл. № 18 (45) Опубликовано: 04.07.2017 Бюл. № 19 (85) Дата начала рассмотрения заявки РСТ на национальной фазе: 20.12.2011 (86) Заявка РСТ: 05 2010/035616 (20.05.2010) (87) Публикация заявки РСТ: УГО 2010/135555 (25.11.2010) Адрес для переписки: 190000, Санкт-Петербург, ул. Малая Морская, 15, офис 5, ВОХ-сервис 1125, ООО "ПАТЕНТИКА" (72) Автор(ы): ГОРДОН-БЕРЕСФОРД, Роланд (ВЕ), ГОССИН, Винциан (ВЕ), ОМСИ, Кристиан (ВЕ), ТЕРЗИК, Андре (05), БЕФАР, Атта (0$) (73) Патентообладатель(и): МАЙО ФАУНДЕЙШН ФО МЕДИКАЛ ЭДЬЮКЕЙШН ЭНД РИСЕРЧ (05), КАРДИОЗ БИОСАЙНСИЗ СА (ВЕ) (56) Список документов, цитированных в отчете о поиске: КАТАЗПМОН .. е а1. ЗТАТЗ- дерепдеп топзе ет6гуопс ет сей ЧИЕегепиайоп шо сагд1лотуосуе$: апа[у$1$ оЁ то[есщат $12па[п? апд {Вегареиис еЁЯсасу оЁ саг1отуосуе ргесоши ед шЕЗ тапзращайоп ш а топзе тоде! оЁ туосаг1а1 шРагсНоп. Сис Кез. 2007 Ост 26; 1010): 910- 918. ЕриБ 2007 5Зер 6 [Найдено 22.12.2014] [он-лайн]. Найдено из (см. прод.) (54) СПОСОБ ОПРЕДЕЛЕНИЯ КАРДИО-ГЕНЕРАТИВНОГО ПОТЕНЦИАЛА КЛЕТОК МЛЕКОПИТАЮЩИХ (57) Реферат: Изобретение ...

КОМПОЗИЦИЯ ДЛЯ ПОДДЕРЖАНИЯ ФУНКЦИИ ТРОМБОЦИТОВ

Изобретение относится к композиции для поддержания функции тромбоцитов, где композиция в качестве активного ингредиента содержит соединение, представленное общей формулой (I), или его фармацевтически приемлемую соль:где соединение, представленное общей формулой (I), представляет собой любое из N-[2-(4-бут-2-инилоксибензолсульфонил)-1-(4-диэтиламинометилфенил)этил]-N-гидроксиформамида и N-{4-[2-(4-бут-2-инилоксибензолсульфонил)-1-(формилгидроксиамино)этил]бензил}метансульфонамида. Изобретение также относится к способу получения тромбоцитов, культуральной среде, препарату крови и к способу поддержания функции тромбоцитов в препарате крови. Технический результат: получена композиция для поддержания функции тромбоцитов посредством специфического ингибирования металлопротеиназной активности ADAM17. 5 н. и 2 з.п. ф-лы, 3 табл., 29 ил., 44 пр.

КОМПОЗИЦИЯ, ВКЛЮЧАЮЩАЯ ЭКЗОСОМУ, ПОЛУЧЕННУЮ ИЗ СТВОЛОВЫХ КЛЕТОК, ДЛЯ ИНДУКЦИИ АДИПОГЕННОЙ ДИФФЕРЕНЦИРОВКИ, РЕГЕНЕРАЦИИ ЖИРОВОЙ ТКАНИ, ОТБЕЛИВАНИЯ КОЖИ ИЛИ КОРРЕКЦИИ МОРЩИН

Изобретение относится к области косметологии и может быть использовано для отбеливания кожи путем снижения синтеза меланина. Предложено применение экзосом, выделенных из пролиферирующих стволовых клеток, полученных из жировой ткани, в качестве активного ингредиента для отбеливания кожи путем снижения синтеза меланина. Изобретение обеспечивает отбеливающий эффект, а также безопасность применения, поскольку экзосомы представляют собой очищенный компонент, не содержащий антибиотики, сыворотки или вредоносные факторы культурального раствора. 2 з.п. ф-лы, 20 ил., 5 табл., 2 пр.

КОМПОЗИЦИЯ, ВКЛЮЧАЮЩАЯ ЭКЗОСОМУ, ПОЛУЧЕННУЮ ИЗ СТВОЛОВЫХ КЛЕТОК, ДЛЯ ИНДУКЦИИ АДИПОГЕННОЙ ДИФФЕРЕНЦИРОВКИ, РЕГЕНЕРАЦИИ ЖИРОВОЙ ТКАНИ, ОТБЕЛИВАНИЯ КОЖИ ИЛИ КОРРЕКЦИИ МОРЩИН

КОМПОЗИЦИЯ ДЛЯ ПОДДЕРЖАНИЯ ФУНКЦИИ ТРОМБОЦИТОВ

... 1. Композиция для поддержания функции тромбоцитов, где композиция в качестве активного ингредиента содержит соединение, представленное приведенной ниже общей формулой (I), или его соль, или его сольват:гдеX представляет собой фениленовую группу;Y представляет собой любой из атома водорода и -(CH)R;гдеm представляет собой любое целое число от 0 до 4; иRпредставляет собой любой изгде Rпредставляет собой любую алкильную группу от C1 до C6, которая может быть замещенной, арильную группу, которая может быть замещенной, и алкоксигруппу от C1 до C6;каждый из Rи Rнезависимо представляет собой любой из атома водорода и алкильной группы от C1 до C6, или Rи Rвместе с соседним атомом азота могут формировать азотсодержащий гетероцикл; иRпредставляет собой любой из атома водорода, алкильной группы от C1 до C6 и алкилсульфонильной группы от C1 до C6; иZ представляет собой любой из атома водорода и алкильной группы от C1 до C6.2. Композиция по п.1, где соединение, представленное общей формулой (I), представляет ...

AMYLOID PEPTIDE INACTIVATING ENZYME TO TREAT ALZHEIMER'S DISEASE

Endocrine precursor cells, pancreatic hormone-expressing cells and methods of production

Cell suspension and use thereof

PCT Patent Application Attorney Docket No. 127630-010501/PCT The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration.

TGFbeta signaling independent naive induced pluripotent stem cells, methods of making and use

Provided is a cocktail of factors for converting/reprogramming non-naïve pluripotent stem cells into TGFβ signaling-independent (TSI) naïve induced pluripotent stem cells (iPSCs). Also provided are methods for reprograming a non-naïve PSC into a TSI naïve iPSC by contacting the cell to be reprogrammed with effective amounts of compounds for a sufficient period of time to reprogram the cell into a TSI naïve iPSC.

심장 조직을 처치하는 세포를 이용하는 조성물 및 방법

... 본 문헌은 심장 인자를 함유하는 조성물, 상기 인자의 존재시 초기 세포를 배양함으로써 세포를 얻는 방법, 및 심장 조직에 대하여 획득한 세포를 투여하는 방법에 관한 것이다.

ENHANCED POSTNATAL ADHERENT CELLS AND USE THEREOF

The present invention relates to an enhanced postnatal adherent cell (ePAC) separated from the placenta and growing by being attached on the surface of a culture container, a manufacturing method thereof, a composition comprising the cell as an active ingredient and a cell medicine. The ePAC is suitable for a medicine by secreting large amounts of proteins, known to be useful for nerve disorders, immune diseases or vessel diseases, thereby being used for developing a cell medicine. COPYRIGHT KIPO 2017 ...

Specification of functional cranial placode derivatives from human pluripotent stem cells

Cranial placodes are embryonic structures essential for sensory and endocrine organ development. The efficient derivation of cranial placodes from human pluripotent stem cells is disclosed where the timed removal of the BMP inhibitor Noggin, a component of the dual-SMAD inhibition strategy of neural induction, triggers placode induction at the expense of CNS fates. Further fate specification at the pre-placode stage enables the selective generation of placode-derived trigeminal ganglia capable of in vivo engraftment, mature lens fibers and anterior pituitary hormone-producing cells that upon transplantation produce hormones including, but not limited to, human growth hormone and adrenocortiocotropic hormone in vivo. Alternatively, anterior pituitary hormone-producing cells are generated in cell culture systems in vitro.

Pluripotent stem cell that can be isolated from body tissue

Objects of the present invention are to provide a method for directly obtaining pluripotent stem cells which do not have tumorigenic property from body tissue and the thus obtained pluripotent stem cells. The present invention relates to SSEA-3 (+) pluripotent stem cells that can be isolated from body tissue.

Use of a Proteolytic Enzyme for the Modification of the Cell Surface of a Stem Cell

The present invention relates to a stem cell and/or a population thereof having a specific profile of cell surface proteins and/or proteoglycans. The present invention also relates to use of a proteolytic enzyme in the modification of the cell surface of a stem cell. The present invention further relates to a method of modifying the cell surface of a stem cell by treatment with a proteolytic enzyme.

Composition for maintaining function of platelets

A composition for maintaining a function of platelets, the composition comprising, as an active ingredient, a compound represented by the following general formula (I) or a salt thereof, or a solvate thereof: wherein X represents a phenylene group; Y represents any one of a hydrogen atom and —(CH 2 ) m R 1 ; wherein m represents an integer of any one of 0 to 4; and R 1 is any one of —NR 5 COR 2 , —NR 5 SO 2 R 2 , and —NR 3 R 4 ; wherein R 2 represents any one of a C1 to C6 alkyl group, an aryl group, a C1 to C6 alkoxy group, and the like; R 3 and R 4 represent a C1 to C6 alkyl group or the like; and R 5 represents any one of a hydrogen atom, a C1 to C6 alkyl group, and the like; and Z represents any one of a hydrogen atom and a C1 to C6 alkyl group.

Method of deriving progenitor cell line

We disclose a method comprising: (a) providing an embryonic stem (ES) cell; and (b) establishing a progenitor cell line from the embryonic stem cell; in which the progenitor cell line is selected based on its ability to self-renew. Preferably, the method selects against somatic cells based on their inability to self-renew. Preferably, the progenitor cell line is derived or established in the absence of co-culture, preferably in the absence of feeder cells, which preferably selects against embryonic stem cells. Optionally, the method comprises (d) deriving a differentiated cell from the progenitor cell line.

Compositions, methods and uses of alpha 1-antitrypsin for early intervention in bone marrow transplantation and treatment of graft versus host disease

Embodiments of the present invention relate to compositions and methods for treatment of subjects in need of or having a bone marrow transplant. Certain embodiments describe compositions and methods for treatment of conditions associated with bone marrow transplantations in a subject, for example, Graft versus Host Disease (GvHD) or bone marrow transplantation rejection. Some embodiments concern early or immediate bone marrow transplantation rejection. Certain embodiments relate to compositions and uses of alpha1-antitrypsin (α1-antitrypsin, AAT) and carboxyterminal peptide derivatives thereof and/or compositions and uses of serine protease inhibitors, immunomodulators or anti-inflammatory agent activity similar to that of AAT.

Genetically modified mesenchymal stem cells expressing alpha-1 antitrypsin (aat)

Genetically modified mesenchymal stem cells can be used as a medicament in the treatment of medical conditions associated with inflammation and/or an unwanted immune response in subjects without an alpha1-antitrypsin (AAT) deficiency. The stem cells include an exogenous nucleic acid, which includes (i) an Alpha-1 antitrypsin (AAT) encoding region operably linked to (ii) a promoter or promoter/enhancer combination.

COMPOSITION FOR EX VIVO CULTURING OF A CORNEAL ENDOTHELIAL CELL AND A METHOD FOR USING THE COMPOSITION

A composition for ex vivo culture of corneal endothelial cell and a method for used the composition are disclosed. The composition or the method is applied to ex vitro culture of the corneal endothelial cell to maintain the corneal endothelial cell in a hexagonal phenotype, and to prevent the corneal endothelial cell from endothelial-mesenchymal transformation (EnMT). Therefore, the corneal endothelial cell can maintain normal phenotype and function and can be used in the further application. The composition at least comprises an effective concentrated matrix metalloproteinase inhibitor (MMPI). 1. A growth regulator for ex vivo culturing of a corneal endothelial cell , at least comprises an effective concentrated matrix metalloproteinase inhibitor (MMPI). The regulator is added in the culture medium to maintain the phenotype and function of corneal endothelial cells and prevent the cells from endothelial-mesenchymal transformating and loss of function.2. The growth regulator as claimed in claim 1 , wherein the concentration of the MMPI is at least 1 uM.3. An inhibitor for preventing an ex vivo cultured corneal endothelial cell form endothelial-mesenchymal transformation claim 1 , at least comprises an effective matrix metalloproteinase inhibitor (MMPI). The inhibitor is added in the culture medium to maintain the phenotype and function of corneal endothelial cells and prevent the cells from endothelial-mesenchymal transformating and loss of function.4. The growth regulator as claimed in claim 3 , wherein the concentration of the MMPI is at least 1 uM.5. A method to prevent a corneal endothelial cells from endothelial-mesenchymal transformation comprising:obtaining the corneal endothelial cell from a donor;incubating the corneal endothelial cell in a corneal endothelial cell culture medium; andadding an effective concentration of matrix metalloproteinase inhibitor in a specific time point.6. The method as claimed in claim 5 , wherein the concentration of the MMPI is ...

Production methods for megakaryocytes and platelets

An object of the present invention is to provide a method of efficiently producing a maturated megakaryocytic cell line from hematopoietic progenitor cells. The present invention provides a method for producing megakaryocytes from hematopoietic progenitor cells, comprising (i) forcibly expressing an apoptosis suppression gene and an oncogene in hematopoietic progenitor cells and culturing the cells, and (ii) arresting forced expression of the apoptosis suppression gene and the oncogene and culturing the hematopoietic progenitor cells.

CONVERSION OF NON-NEURONAL CELLS INTO NEURONS

The subject invention concerns materials and methods for conversion of non-neuronal cells into neurons. The subject invention also concerns methods for screening drugs and other compounds for activity in treating Alzheimer's disease using neuronal cells produced using the subject invention. The subject invention also concerns neuronal cells that have been produced using the methods of the invention. The subject invention also concerns methods for evaluating therapeutic treatment for efficacy in a person or animal having Alzheimer's disease or other neurodegenerative diseases. The subject invention also concerns methods of treating Alzheimer's disease or other neurodegenerative diseases or conditions in a person or animal. 1. A neuronal cell produced by conversion of a non-neuronal cell to a neuronal cell by:i) incorporating into said non-neuronal cell a polynucleotide of polypyrimidine-tract-binding (PTB) protein short hairpin RNA (shRNA) and a polynucleotide encoding a selectable marker;ii) selecting for a cell expressing said selectable marker;iii) culturing said selected cell in a neural induction medium comprising a ROCK inhibitor for a sufficient period of time to induce neuronal development, whereby said selected cell is converted into a neuronal cell.2. The neuronal cell according to claim 1 , wherein said polynucleotide is incorporated into said non-neuronal cell using a virus or viral vector.3. The neuronal cell according to claim 2 , wherein said virus or vector is adenovirus claim 2 , AAV claim 2 , retrovirus claim 2 , or lentivirus.4. The neuronal cell according to claim 1 , wherein said cell is a human cell.5. The neuronal cell according to claim 1 , wherein said non-neuronal cell is a fibroblast cell.6. The neuronal cell according to claim 1 , wherein said neuronal cell expresses one or more of tubulin claim 1 , MAP2 claim 1 , synapsin claim 1 , or NeuN.7. A method for screening a drug or compound for activity in treating or inhibiting Alzheimer's ...

GUIDED DIFFERENTIATION OF INDUCED PLURIPOTENT STEM CELLS

This document provides methods and materials related to making and using differentiated induced pluripotent stem cells. For example, methods and materials for making differentiated induced pluripotent stem cells (e.g., insulin-producing cells) that do not form cancer cells within a mammal (e.g., a human), cells that underwent guided differentiation from induced pluripotent stem cells, compositions containing cells that underwent guided differentiation from induced pluripotent stem cells, and methods for using cells that underwent guided differentiation from induced pluripotent stem cells (e.g., methods for using such cells to treat diabetes or to repair cardiovascular tissue) are provided. 1. A population of differentiated cells obtained from induced pluripotent stem cells , wherein the cells of said population lack integrated viral nucleic acid encoding a stemness factor , wherein implantation of said population of differentiated cells into a mammal does not result in cancer cell formation , and wherein said cells were obtained using a culture comprising an enzyme.2. The population of differentiated cells of claim 1 , wherein said cells are human cells.3. The population of differentiated cells of claim 1 , wherein said cells lack exogenous nucleic acid.4. The population of differentiated cells of claim 1 , wherein said enzyme is trypsin.5. A method for obtaining a population of differentiated cells obtained from induced pluripotent stem cells claim 1 , wherein said method comprises:(a) exposing somatic cells to one or more non-integrating viral vectors that direct the expression of one or more stemness factors to produce induced pluripotent stem cells from said somatic cells, and(b) exposing said induced pluripotent stem cells to one or more differentiation factors in the presence of an enzyme to produce a population of cells more specialized than said induced pluripotent stem cells.6. The method of claim 5 , wherein said somatic cells are keratinocytes.7. The ...

Methods for production of platelets from pluripotent stem cells and compositions thereof

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.

Method for cell culture

A method for stem or progenitor cell culture. More precisely, the invention relates to a method for cell culture using one or more IαI (inter-alpha trypsin inhibitor or inter-alpha inhibitor) protein(s) or part(s) thereof as a component in a cell culture media or a coating on a cell culture surface material. Furthermore the invention relates to a cell culture media and a cell culture coating/matrix provided with one or more IαI proteins(s) or part(s) thereof.

METHODS FOR PRODUCTION OF PLATELETS FROM PLURIPOTENT STEM CELLS AND COMPOSITIONS THEREOF

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells. 1. A pharmaceutical preparation that is suitable for use in a human patient comprising at least 10platelets , wherein the preparation is substantially free of leukocytes and wherein substantially all of the platelets are functional.26.-. (canceled)7. A bioreactor having weakly adherent or non-adherent megakaryocytes that produce functional platelets without feeder cells.8. A composition , a cryopreserved composition , or a bank comprising MLPs.913.-. (canceled)14. A method for producing platelets from megakaryocytes comprising:(a) providing a non-adherent culture of megakaryocytes;(b) contacting the megakaryocytes with (i) TPO or a TPO agonist, (ii) hematopoietic expansion medium and optionally TPO or a TPO agonist, SCF, IL-6 and IL-9, or (iii) hematopoietic expansion medium and optionally TPO or a TPO agonist, SCF, and IL-11 to cause the formation of proplatelets in culture, wherein the proplatelets release platelets; and(c) isolating the platelets.1535.-. (canceled)36. The method of claim 14 , wherein the megakaryocytes are generated by:(a) culturing pluripotent stem cells to form hemogenic endothelial cells (PVE-HE);(b) culturing the hemogenic endothelial cells to form MLPs; and(c) culturing the MLPs to form megakaryocytes.3750.-. (canceled)51. A method of treating a patient in need of platelet transfusion claim 1 , comprising administering to the patient the ...

REGULATION OF DIFFERENTIATION INTO DOPAMINERGIC NEURONS BY METALLOPROTEASE

The present invention relates to a method for regulating the differentiation of neural stem cells or neural progenitor cells into dopaminergic neural cells, the method comprising increasing or inhibiting the activity of ADAM17 and/or ADAM10 in neural stem cells or neural progenitor cells, and to the use of an activator or inhibitor of ADMA17 and/or ADAM10. 1. A method for regulating the differentiation of neural stem cells or neural progenitor cells into dopaminergic neural cells , the method comprising increasing or inhibiting the activity of ADAM17 and/or ADAM10 in neural stem cells or neural progenitor cells.2. The method of claim 1 , wherein the activity of ADAM17 is increased using one or more selected from the group consisting of 12-HPETE (12-hydroperoxy-5Z claim 1 ,8Z claim 1 ,10E claim 1 ,14Z-eicosatetraenoic acid) claim 1 , bortezomib (Millennium Pharmaceuticals claim 1 , USA) claim 1 , Furin claim 1 , GM-CSF claim 1 , N-formyl-L-methionyl-phenylalanine claim 1 , and PMA (phorbol 12-myristate-13-acetate).3. The method of claim 1 , wherein the activity of ADAM17 is inhibited using one or more selected from the group consisting of TAPI-1 (CHNO) claim 1 , TAPI-2 (CHNO) claim 1 , GW3333 (CHNO) claim 1 , GW280264X (hydroxamate) claim 1 , siRNA claim 1 , and antisense RNA.4. The method of claim 1 , wherein the activity of ADAM10 is increased using one or more selected from the group consisting of 5α-dihydrotestosterone claim 1 , donepezil claim 1 , EGF (epidermal growth factor) claim 1 , PMA (phorbol-12 myristate 13-acetate) claim 1 , and thyrotropin.5. The method of claim 1 , wherein the activity of ADAM10 is inhibited using one or more selected from the group consisting of TAPI-1 (CHNO) claim 1 , TAPI-2(CHNO) claim 1 , GI254023X (((2R claim 1 ,3S)-3-(formyl-hydroxyamino)-2-(3-phenyl-1-propyl)butanoic acid)[(1S)-2 claim 1 ,2-dimethyl-1-methylcarbamoyl-1-propyl]amide) claim 1 , GM6001 ((2S)—N4-hydroxy-N1-[(1S)-1-(1H-indol-3-ylmethyl)-2-(methylamino)-2-oxoethyl]-2 ...

CELL POPULATION INCLUDING MESENCHYMAL STEM CELLS AND PRODUCTION METHOD THEREFOR, AND PHARMACEUTICAL COMPOSITION

An object of the present invention is to provide a cell population comprising mesenchymal cells having low cell aggregability, which is useful for intravenous administration of a cell preparation, and a method for producing the same, and a pharmaceutical composition comprising the cell population. Also provided are methods for producing a cell population comprising mesenchymal stem cells, the method comprising obtaining a cell population having the following cell characteristics (the cell population satisfies the relative expression level of COL11A1 gene to the expression level of SDHA gene of 6.0 or less and the relative expression level of COL16A1 gene to the expression level of SDHA gene of 1.5 or less). 1. A method for producing a cell population comprising mesenchymal stem cells , the method comprisingobtaining a cell population having the following cell characteristics: the cell population satisfies the relative expression level of COL11A1 gene to the expression level of SDHA gene of 6.0 or less and the relative expression level of COL16A1 gene to the expression level of SDHA gene of 1.5 or less.2. The method of claim 1 , wherein the step of obtaining comprisestreating a sample comprising an extracellular matrix layer collected from a fetal appendage with an enzyme solution to release the mesenchymal stem cells contained in the extracellular matrix layer; andidentifying the cell population having the cell characteristics from the released mesenchymal stem cells.3. The method of claim 2 , wherein the step of obtaining further comprises selectively separating the identified cell population.4. The method of claim 2 , wherein the enzyme solution comprises a collagenase claim 2 , a metalloproteinase claim 2 , or both.5. The method of further comprising culturing or cryopreserving the cell population.6. The method of claim 1 , wherein the cell population has a low cell aggregability.7. The method of claim 1 , wherein the cell population can be cultured up to 40 days ...

PRODUCTION OF YOGURT

The present invention is in the field of fermentation, and pertains to a method for preparing yogurt by fermentation, comprising the steps of providing a fermentation starter culture comprising a selected microorganism in a suitable culture medium, adding a potato protein protease inhibitor to the culture medium, culturing the microorganism, and harvesting the yogurt. This method has the advantage that the lag time of fermentation can be reduced by the addition of relatively low amounts of potato protein protease inhibitor. It has the additional advantage that the potato protein protease inhibitor allows for application of the present method over a wide pH range and wide temperature range including pasteurization, is filter sterilizable and that potato protease inhibitor protein is non-allergenic. 1. A method for preparing yogurt , comprising the steps of providing a fermentation starter culture comprising a selected microorganism in a suitable culture medium , adding a plant protein protease inhibitor to the culture medium , culturing the microorganism in the culture medium , and harvesting the yogurt.2. A method for fermenting yogurt according to claim 1 , wherein growth of the microorganism is peptide-limited.3. A method according to claim 1 , wherein the plant protein protease inhibitor is a potato protein protease inhibitor.4. A method according to claim 1 , wherein the microorganism is chosen from the group of lactic acid bacteria and yeasts.5Streptococcus, Lactobacillus, Lactococcus, Carnobacterium, LeuconostocPediococcus,Bifidobacteriales.. A method according to claim 1 , wherein the microorganism is chosen from the group of and or from the order of6. A method according to claim 1 , wherein the culture medium comprises milk.7. A method according to claim 1 , wherein the potato protein protease inhibitor is present in the culture medium in an amount between 5 g/l and 0.001 g/l.8. A method according to claim 1 , wherein the pH of the culture medium is between ...

A METHOD FOR THE REGENERATION AND DIFFERENTIATION OF HUMAN PERINEPHRIC FAT DERIVED MESENCHYMAL STROMAL CELLS INTO ASTROGLIAL, RENAL, NEURONAL AND PANCREATIC PROGENITOR CELLS

The embodiments of the present invention provide a method for the regeneration and differentiation of human perinephric fat derived mesenchymal stromal cells (hPNF-MSCs) into astroglial, renal, neuronal progenitor cells and pancreatic islet cells. The hPNF-MSCs are advantageous over bone marrow or other stem cell source due to the abundance and ease of isolation. The perinephric fat derived stromal cells are used in the treatment of neurodegenerative disorder, renal disorder and pancreatic malfunction. The method for the regeneration and differentiation of hPNF-MSCs includes isolation of the human perinephric fat derived stromal cells followed by expansion of the stem cells and cryopreservation. The cells are characterized by morphological evaluation of hPNF-MSC, growth kinetics, immunophenotypic evaluation, differentiation potential analysis, karyotyping and secretome profiling (cytokines, chemokines, and growth factors). The stem cells are differentiated into astroglial cells, neural cells renal cells and pancreatic islet cells. 1. A method for isolating human perinephric fat derived stem/stromal cells (hPNF-MSCs) , for propagation , regeneration and differentiation into astroglial cells , renal cells , neuronal cells and islet cells , the method comprises the steps of:collecting the perinephric fat tissue from a volunteer or a kidney patient with a prior informed consent;processing the perinephric fat tissue in a sterile environment, and wherein the sterile environment comprises Current Good Manufacturing Practices (cGMP) compliant clean room with a biosafety cabinet;isolating the mesenchymal stem/stromal cells from the perinephric fat tissues using a plurality of methods, and wherein the plurality of methods are a mechanical method and an enzymatic method;washing the tissue sample in a 1× Dulbecco's phosphate Buffered Saline (DPBS) solution comprising of antibiotics at a concentration of 2× concentration, and wherein the tissue sample is washed for a plurality ...

AGENTS AND METHODS FOR TREATING AND PREVENTING SEBORRHEIC KERATOSIS

Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex vivo. Also provided herein are screening assays using cultured seborrheic keratosis cells and methods for treating seborrheic keratosis in a subject. 1. A method for inducing apoptosis in a seborrheic keratosis cell , the method comprising contacting a seborrheic keratosis cell with a composition comprising an effective amount of an ATP-competitive Akt inhibitor , thereby inducing apoptosis in the cell.2. The method of claim 1 , wherein the effective amount of the ATP-competitive Akt inhibitor or the composition does not substantially affect the survival of normal keratinocytes.34.-. (canceled)5. The method of claim 1 , wherein the ATP-competitive Akt inhibitor is A-443654 claim 1 , AT7867 claim 1 , or GSK690693.6. A method for treating a seborrheic keratosis in a subject claim 1 , the method comprising administering a composition comprising a therapeutically effective amount of an ATP-competitive Akt inhibitor to a subject having seborrheic keratosis.7. The method of claim 6 , wherein the composition is applied topically or administered systemically.8. The method of claim 6 , wherein the therapeutically effective amount of the ATP-competitive Akt inhibitor or the composition does not substantially affect the survival or normal keratinocytes.9. The method of claim 6 , wherein the composition further comprises a pharmaceutically acceptable carrier.1012.-. (canceled)13. The method of claim 6 , further comprising a step of diagnosing the subject with a seborrheic keratosis.14. The method of claim 6 , wherein the ATP-competitive Akt inhibitor is A-443654 claim 6 , AT7867 claim 6 , or GSK690693. This application is a Continuation of U.S. application Ser. No. 16/029,892, filed Jul. 9, 2018, which is a continuation of U.S. application Ser. No. 15/482,899 filed on Apr. 10, 2017, which is a continuation of U.S. application Ser. No. 15/254,344 (now U.S. Pat. No. 9,658,210), filed ...

METHODS AND MATERIALS FOR USING FIBRIN SUPPORTS FOR RETINAL PIGMENT EPITHELIUM TRANSPLANTATION

This document provides methods and materials for performing retinal pigment epithelium transplantation. For example, methods and materials for using fibrin supports for retinal pigment epithelium transplantation are provided. 118-. (canceled)19. A retinal implant comprising (a) a retinal pigment epithelium monolayer having an apical surface and a basal surface , and (b) a fibrin hydrogel layer attached to said basal surface of said monolayer.20. The implant of claim 19 , wherein said fibrin hydrogel layer is from about 20 μm to about 400 μm thick.21. The implant of claim 19 , wherein said implant comprises plasminogen.22. The implant of claim 19 , wherein said implant comprises from about 0.1 U of plasminogen per mL to about 40 U of plasminogen per mL or from about 0.001 U of plasminogen per mL to about 40 U of plasminogen per mL.23. The implant of claim 19 , wherein said fibrin hydrogel layer comprises a coating.24. The implant of claim 23 , wherein said coating comprises basement membrane proteins claim 23 , matrigel claim 23 , or geltrex.25. A method for making a retinal implant claim 23 , wherein said method comprises:(a) obtaining a fibrin hydrogel layer,(b) coating a surface of said fibrin hydrogel layer with an agent, and(c) forming a retinal pigment epithelium monolayer having an apical surface and a basal surface on said coating, wherein said basal surface is closer to said fibrin hydrogel layer than said apical surface.26. The method of claim 25 , wherein said fibrin hydrogel layer is from about 20 μm to about 400 μm thick.27. The method of claim 25 , wherein said fibrin hydrogel layer comprises from about 20 mg of fibrinogen per mL to about 80 mg of fibrinogen per mL.28. The method of claim 25 , wherein said fibrin hydrogel layer comprises from about 2 U of thrombin per mL to about 1500 U of thrombin per mL.29. The method of claim 25 , wherein said fibrin hydrogel layer comprises from about 0.1 U of plasminogen per mL to about 40 U of plasminogen per mL ...

Cell Differentiation Marker and its Uses

The use of Dub3 protein, a nucleic acid molecule coding for the protein, or an inhibitor of the activity and/or of the expression of the protein for modulating cell differentiation. 1. A method for modulating cell differentiation comprising the administration to a determined cell:Dub3 protein, said protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any variant thereof having at least 43% identity with said amino acid sequence SEQ ID NO: 1, and having ubiquitin hydrolase activity ora nucleic acid molecule coding for said protein or said variant thereof, oran inhibitor of the activity, i.e. the ubiquitin hydrolase activity and/or of the expression of said protein or said variant thereof.2. The method according to claim 1 , for modulating totipotent or pluripotent cell differentiation.3. A method for inducing dedifferentiation of differentiated cells claim 1 , the cells obtained from the dedifferentiation of differentiated cells being iPS cells claim 1 , the method comprising a step of administering to a differentiated cellsDub3 protein, said protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any variant thereof having at least 43% identity with said amino acid sequence SEQ ID NO: 1, ora nucleic acid molecule coding for said protein or said variant thereof.4. The method according to for inducing dedifferentiation of differentiated cells claim 3 , wherein said cells Dub3 protein or said nucleic acid molecule coding for said protein are associated with at least an Oct family member protein and a Sox family member protein.5. The method according to claim 3 , wherein said Dub3 protein is expressed in said iPS cells at a level corresponding to at least 2 fold lower than the expression of said Dub3 protein in totipotent cell.6. A method for inducing a spontaneous differentiation of totipotent or pluripotent cells claim 3 , comprising the administration to a determined cell of an inhibitor of the activity and/or of the ...

METHODS OF PRODUCING RPE CELLS AND COMPOSITIONS OF RPE CELLS

The present invention provides improved methods For producing RPE cells from human embryonic stem cells or from other human pluripotent stem cells. The invention also relates to human retinal pigmented epithelial cells derived from human embryonic stem cells or other human multipotent or pluripotent stem cells. hRPE cells derived from embryonic stem cells are molecularly distinct from adult and fetal-derived RPE cells, and are also distinct from embryonic stem cells. The hRPE cells described herein are useful for treating retinal degenerative diseases. 1. A method of producing a substantially purified culture of retinal pigment epithelial (RPE) cells , comprisinga) providing human embryonic stem cells;b) culturing the human embryonic stem cells as embryoid bodies in nutrient rich, low protein medium;c) culturing the embryoid bodies as an adherent culture in nutrient rich, low protein medium;d) culturing the adherent culture of cells of (c) in nutrient rich, low protein medium, which medium does not contain serum free B-27 supplement;e) culturing the cells of (d) in medium capable of supporting growth of high-density somatic cell culture, whereby RPE cells appear in the culture of cellsf) contacting the culture of (e) with an enzyme;g) selecting the RPE cells from the culture and transferring the RPE cells to a separate culture containing medium supplemented with a growth factor to produce an enriched culture of RPE cells; and (i) a substantially purified culture of RPE cells, and/or', '(ii) mature RPE cells., 'h) propagating the enriched culture on RPE cells to produce'}237-. (canceled)38. A method of treating or preventing a condition characterized by retinal degeneration claim 1 , comprising administering to a subject in need thereof an effective amount of a preparation comprising RPE cells claim 1 , which RPE cells are derived from human pluripotent stem cells according to the method of .3952-. (canceled)53. A method of producing a substantially purified culture ...

Cell Suspension and Use Thereof

The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration. 1. A cell suspension for use in an epithelium-related procedure , comprising:a population of cells including viable and functioning cells derived from an epithelial tissue sample;wherein the population of cells, prior to being used in an epithelium-related procedure, are exposed to a condition selected from the group consisting of stress and an exogenous agent;wherein the population of cells, when used in said epithelium-related procedure and applied to a recipient site, promote treatment, healing, reconstructing, resurfacing, repigmentation and/or regeneration of epithelial tissues.2. The cell suspension of claim 1 , wherein the viable and functioning cells include differentiated cells and cells capable of dividing or differentiating.3. The cell suspension of claim 1 , wherein the epithelial tissue sample is obtained from one or more of skin epithelium claim 1 , respiratory epithelium claim 1 , vascular epithelium claim 1 , corneal epithelium claim 1 , and glandular epithelium.4. The cell suspension of claim 1 , wherein when the epithelial tissue sample is skin tissue ...

METHODS OF PRODUCING RPE CELLS AND COMPOSITIONS OF RPE CELLS

The present invention provides improved methods for producing RPE cells from human embryonic stem cells or from other human pluripotent stem cells. The invention also relates to human retinal pigmented epithelial cells derived from human embryonic stem cells or other human multipotent or pluripotent stem cells. hRPE cells derived from embryonic stem cells are molecularly distinct from adult and fetal-derived RPE cells, and are also distinct from embryonic stem cells. The hRPE cells described herein are useful for treating retinal degenerative diseases. 1. A method of producing a substantially purified culture of retinal pigment epithelial (RPE) cells , comprisinga) providing human pluripotent stem cells;b) culturing the human pluripotent stem cells as embryoid bodies or aggregates in nutrient rich, low protein medium that comprises less than 5% protein;c) culturing the embryoid bodies or aggregates as an adherent culture in nutrient rich, low protein medium that comprises less than 5% protein;d) culturing the cells of (c) in medium capable of supporting growth of high-density somatic cell culture, whereby RPE cells appear in the culture of cells;e) contacting the culture of (d) with an enzyme;f) selecting the RPE cells from the culture and transferring the RPE cells to a separate culture containing medium supplemented with a growth factor to produce an enriched culture of RPE cells; andh) propagating the enriched culture on RPE cells to produce a substantially purified culture of RPE cells.2. The method of claim 1 , wherein the RPE cells are further cultured to produce a culture of mature RPE cells.36-. (canceled)7. The method of claim 1 , wherein the cells of (b) are cultured for 7-14 days.8. (canceled)9. The method of claim 1 , wherein the cells of (c) are cultured for 7-10 days.10. The method of claim 1 , wherein the cells of (d) are cultured for 14-21 days.1115-. (canceled)16. The method of claim 1 , wherein the growth factor of (f) is selected from the group ...

METHODS OF USING PDX1-POSITIVE PANCREATIC ENDODERM CELLS AND ENDOCRINE PRECURSOR CELLS

Disclosed herein are cell cultures and enriched cell populations of endocrine precursor cells, immature pancreatic hormone-expressing cells and mature pancreatic hormone-expressing cells. Also disclosed herein are methods of producing such cell cultures and cell populations.

2d organoid for infection and culture of human diarrhea virus, and use of said 2d organoid

A 2D organoid for infection and growth culture of human diarrhea virus, obtained by: (1) obtaining a 3D organoid by three-dimensionally culturing a human intestinal epithelial stem cell, a human intestinal epithelial cell, or a tissue containing at least any of those cells on an extracellular matrix; and (2) dispersing the 3D organoid to prepare a single cell, and monolayer-culturing the single cell on an extracellular matrix to obtain the 2D organoid having a monolayer structure in which epithelial cells contain differentiated trophoblastic cells and goblet cells and configured as human intestinal lumen having a monolayer structure.

CELL SUSPENSION AND USE THEREOF

The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration. 149-. (canceled)50. A method of preparing a cell suspension for tissue regeneration , the method comprising:collecting a population of cells from a skin tissue sample; andexposing the population of cells to platelet-enriched plasma.51. The method of claim 50 , wherein the skin tissue sample comprises an autologous epithelial tissue sample.52. The method of claim 50 , wherein the population of cells comprises viable and functioning cells from the skin tissue sample.53. The method of claim 52 , wherein the population of cells comprises keratinocytes claim 52 , melanocytes claim 52 , and fibroblasts.54. The method of claim 50 , wherein collecting the population of cells comprises exposing the skin tissue sample to an enzyme to dissociate cellular layers in the skin tissue sample.55. The method of claim 54 , wherein the enzyme comprises at least one enzyme selected from the group consisting of trypsin claim 54 , trypsin-EDTA claim 54 , dispase claim 54 , collagenase claim 54 , thermolysin claim 54 , pronase claim 54 , hyaluronidase claim 54 , elastase claim 54 , papain claim ...

CELL SUSPENSION AND USE THEREOF

The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration. 149-. (canceled)50. A method for treating a chronic wound in skin of a patient , comprising:preparing an autologous cell suspension without in vitro culturing; andadministering the cell suspension to the chronic wound of the patient to promote tissue regeneration at the chronic wound.51. The method of claim 50 , wherein the chronic wound is a chronic ulcer.52. The method of claim 51 , wherein the chronic ulcer is a diabetic ulcer.53. The method of claim 51 , wherein the chronic ulcer is at least one selected from the group consisting of a venous ulcer claim 51 , an arterial ulcer claim 51 , and a mixed arterio-venous ulcer.54. The method of claim 50 , wherein the chronic wound is at least one selected from the group consisting of a neuropathic ulcer and a pressure sore.55. The method of claim 50 , wherein preparing the autologous cell suspension comprises obtaining a skin tissue sample from the patient claim 50 , collecting a population of cells from the skin tissue sample claim 50 , and mixing the population of cells with a nutrient solution to create the autologous ...

METHOD FOR PRODUCING ADULT LIVER PROGENITOR CELLS

Novel adult liver progenitor cells (called H2Stem Cells) have been have been characterized on the basis of a series of biological activities and markers. Methods for producing H2Stem Cells allow providing such cells in the form of adherent cells and three-dimensional cell clusters in suspension that can be differentiated into cells having strong liver-specific activities and/or that can be used for treating liver diseases or for evaluating the efficacy, the metabolism, and/or toxicity of a compound. 146-. (canceled)47. A method of treating a condition , the method comprising administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of: (a) positive for α-smooth muscle actin (ASMA), albumin (ALB), and CD140b, and', '(b) negative for Sushi domain containing protein 2 (SUSD2) and Cytokeratin-19 (CK-19), or, '(i) a human adult liver progenitor cell wherein said cell is measured(ii) an isolated cell population, wherein at least 60% of the cells of the population are cells of (i),wherein the condition may benefit from engraftment of said cells in a human tissue of the subject.48. The method of claim 47 , wherein said composition is administered to said subject by intrahepatic claim 47 , intrasplenic claim 47 , or intravenous administration.49. The method of claim 47 , wherein said cell of (i) is further measured positive for:(a) At least one hepatic marker selected from the group consisting of albumin, HNF-3B, HNF-4, CYP1A2, CYP2C9, CYP2E1 and CYP3A4;(b) At least one mesenchymal marker selected from the group consisting of Vimentin, CD90, CD73, CD44, and CD29;(c) At least one liver-specific activity selected from the group consisting of urea secretion, bilirubin conjugation, alpha-1-antitrypsin secretion, and CYP3A4 activity;(d) At least one marker selected from the group consisting of ATP2B4, ITGA3, TFRC, SLC3A2, CD59, ITGB5, CD151, ICAM1, ANPEP, CD46, and CD81; and(e) At least one marker selected from the group ...

EX VIVO METHODS FOR PRODUCING A T CELL THERAPEUTIC AND RELATED COMPOSITIONS AND METHODS

Provided herein are methods for ex vivo expansion of a T cells including tumor-reactive T cells, and compositions containing such T cells. Also provided are methods for treating diseases and conditions such as cancer using compositions of the present disclosure. 1. A method for manufacturing tumor-reactive T cells , the method comprising: (a) incubating a population of cells comprising T cells from a biological sample obtained from a subject that has a tumor with a first T cell stimulatory agent(s) under conditions to stimulate expansion of T cells of the population to produce a population of stimulated T cells;', '(b) co-culturing the population of stimulated T cells in the presence of antigen presenting cells (APCs) under conditions in which the APCs have been induced to present one or more peptides from a tumor-associated antigen from the subject, thereby generating a population containing T cells comprising tumor reactive T cells;', '(d) enriching from the co-culture the population of tumor reactive T cells reactive to the one or more peptides, wherein said tumor-reactive T cells comprise an endogenous TCR that is reactive to a tumor-associated antigen, thereby producing a population of T cells enriched for tumor-reactive T cells; and', '(d) incubating the population of T cells enriched in tumor reactive T cells with a second T cell stimulatory agent(s) under conditions to stimulate expansion of T cells in the population; and', 'wherein one or more steps of the culturing is carried out in the presence of at least one T cell adjuvant that is an apoptosis inhibitor that inhibits caspase activation or activity; and, '(1) culturing T cells by a process that comprises(2) harvesting cells produced by the method to produce a composition of expanded T cells enriched in tumor reactive T cells.2. The method of claim 1 , wherein the first or second T cell stimulatory agent(s) comprises the presence of one or more recombinant cytokines claim 1 , wherein the incubating in ...

METHOD FOR PREPARING DIFFERENTIATION-INDUCED CELLS

A method for preparing differentiation-induced cells from embryoid bodies derived from pluripotent stem cells is provided. The method includes adding a protease to embryoid bodies. The protease disperses the embryoid bodies, and the protease has an enzyme activity in the range from 0.3 to 4.0 recombinant protease activity unit (rPU)/ml. 1. A method for preparing differentiation-induced cells from embryoid bodies derived from pluripotent stem cells , the method comprising:adding a protease to embryoid bodies such that the protease disperses the embryoid bodies, the protease having an enzyme activity in a range from 0.3 to 4.0 recombinant protease activity unit (rPU)/ml.2. The method according to claim 1 , wherein the protease has an enzyme activity of at least 0.45 rPU/ml.3. The method according to claim 1 , wherein the protease has an enzyme activity from 0.9 to 1.2 rPU/ml.4. The method according to claim 1 , wherein the protease is xeno free.5. The method according to claim 1 , wherein the protease is TrypLE (registered trademark) Select.6. The method according to claim 1 , further comprising treating the embryoid body with collagenase.7. The method according to claim 1 , wherein the pluripotent stem cells are induced pluripotent stem (iPS) cells.8. The method according to claim 1 , wherein the pluripotent stem cells are human cells.9. The method according to claim 1 , wherein the pluripotent stem cells are feeder-free cell lines.10. The method according to claim 1 , wherein the differentiation-induced cell is a cardiomyocyte.11. The method according to claim 1 , wherein a cell population having a troponin positive rate of 50 to 90% is obtained.12. The method according to claim 1 , wherein the dispersed embryoid bodies have a diameter of 10 μm or more.13. The method according to claim 1 , further comprising adhering the dispersed embryoid bodies to a culture substrate.14. A method of improving adhesion of embryoid bodies to a culture substrate comprising:adding a ...

METHOD FOR CULTURING PRIMARY CELLS FROM SOLID TUMOR OF LUNG CANCER AND PRIMARY TUMOR CELLS FROM PLEURAL EFFUSION OF LUNG CANCER AND AUXILIARY REAGENTS

A method for culturing primary cells from solid tumor tissues of lung cancer and primary tumor cells from pleural effusion of lung cancer, and the auxiliary reagents. A method for culturing primary cells from solid tumor tissues of lung cancer and primary tumor cells from pleural effusion of lung cancer and the auxiliary reagents. The core of the technology is as follows: (1) solid tumor of lung cancer are treated by using a mild cell dissociating reagent, and lung cancer cells in pleural effusion are dissociated by a mild method, ensuring the vitality of cancer cells to the greatest extent; (2) a special serum-free medium is prepared, and tumor cells derived from solid tumor tissues of lung cancer are cultured in vitro by using a suspension culture system, ensuring the normal amplification of the cancer cells while eliminating the interference of normal cells to the greatest extent. 122-. (canceled)23. A medium for culturing primary cells of lung cancer , comprising a dual-antibody P/S , HEPES , a non-essential amino acid solution , GlutaMax , human recombinant protein EGF , human recombinant protein bFGF , human recombinant protein MSP , cortisol , B27 , ITS-X , Y-27632 and an Advanced DMEM/F12 medium; wherein the final concentration of penicillin in the dual-antibody P/S is 100-200 U/mL; the final concentration of streptomycin in the dual-antibody P/S is 100-200 μg/mL the final concentration of the HEPES is 8-12 mM; the final concentration of the non-essential amino acid solution is 0.8-1.2% (volume percentage); the final concentration of the GlutaMax is 0.8-1.2% (volume percentage); the final concentration of the human recombinant protein EGF is 10-100 ng/mL; the final concentration of the human recombinant protein bFGF is 10-50 ng/mL; the final concentration of the human recombinant protein MSP is 5-25 ng/mL; the final concentration of the cortisol is 20-50 ng/mL; the final concentration of the B27 is 1.5-2.5% (volume percentage); the final concentration of the ...

INHIBITION OF PROTEIN DEGRADATION FOR IMPROVED PRODUCTION

Disclosed herein are methods and compositions useful for evaluating, selecting, identifying, or making a cell or cell line that has improved production capacity for generating higher yields of products and/or improved capacity to produce higher quality products. Products, as described herein, can include a polypeptide that is endogenously expressed by the cell, a recombinant polypeptide that is not endogenously expressed, or a non-naturally occurring recombinant polypeptide. The methods described herein include modulating, e.g., inhibiting, the protein degradation pathway by using a proteasome inhibitor, an ER-associated degradation (ERAD) inhibitor, or a ubiquitin pathway inhibitor. 1. A method of evaluating a cell or a progeny cell or population of progeny cells for production of a product , comprising:a) providing a cell;b) contacting the cell (or progeny of the cell) concurrently or sequentially with a first and a second inhibitor of protein degradation, wherein the inhibitors of protein degradation are selected from a proteasome inhibitor, a ubiquitin pathway inhibitor, or an endoplasmic reticulum associated degradation (ERAD) inhibitor; andc) evaluating the effect of the first and second inhibitors of protein degradation on one or more parameters related to cell function, in the cell (or progeny of the cell) by comparing a value for the effect of the inhibitors with a reference value;thereby evaluating a cell or a progeny cell or population of progeny cells.2. The method of claim 1 , further comprising as a part of step (b) claim 1 , culturing the cell (or progeny of the cell) in contact with the inhibitor of protein degradation to provide a population of cultured cells or a population of progeny cells.3. The method of claim 1 , comprising responsive to the value meeting or exceeding the reference value claim 1 , identifying claim 1 , classifying claim 1 , or selecting claim 1 , a cell or a progeny cell or population of progeny cells for establishing a culture ...

EX VIVO MATURATION OF ISLET CELLS

The invention relates to methods for promoting maturation of islet cells from pre-weaned mammals for the purpose of optimizing the islets for their use as donor tissue for xenotransplantation. The method of the invention removes the pancreas from donor animals and reduces the pancreas tissue to fragments that are greater than the size of an intact islet while retaining islets in their whole, insulin-producing condition. The method of the invention also serially cultures the digested tissue in novel maturation media that enhance the glucose responsiveness of the cultured islets, and selects islets that are sufficiently glucose-responsive for use in transplantation procedures. 1. A method for promoting the maturation of islet cells from a pre-weaned mammal , comprising:a) removing a pancreas from a pre-weaned mammal;b) reducing the pancreas tissue to fragments that are greater than the size of an intact islet while retaining islets in their whole, insulin-producing condition;c) digesting the tissue fragments in a protease solution such that any exocrine tissue that surrounds the islets is only partially separated from the islets;d) culturing the digested tissue in a first maturation medium;e) culturing the digested tissue in a second maturation medium;f) determining the glucose responsiveness of the cultured islets; andg) selecting glucose-responsive islets.2. The method of claim 1 , wherein the pre-weaned mammal is a piglet.3. The method of claim 1 , wherein the first maturation medium comprises:a) an undefined component of animal origin; b) an antioxidant compound; c) a component that enhances glucose-dependent insulin secretion by beta cells; d) a derivative of vitamin B; f) a derivative of vitamin E; g) heparin; h) a basic pancreatic trypsin inhibitor; i) a serine protease inhibitor; and j) a deoxyribonuclease4. The method of claim 1 , wherein the second maturation medium comprises:a) an undefined component of animal origin; b) an antioxidant compound; c) a ...

METHODS FOR PRODUCTION OF PLATELETS FROM PLURIPOTENT STEM CELLS AND COMPOSITIONS THEREOF

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells. 1. A pharmaceutical preparation that is suitable for use in a human patient comprising at least 10platelets , wherein the preparation is substantially free of leukocytes and wherein substantially all of the platelets are functional , ora bioreactor having weakly adherent or non-adherent megakaryocytes that produce functional platelets without feeder cells, or{'sup': '9', 'a composition comprising at least 10MLPs, or'}{'sup': 9', '14', '9', '10', '11', '12', '13', '14, 'a cryopreserved composition comprising MLPs, optionally comprising comprising 10to 10MLPs, further optionally 10, 10, 10, 10, 10or 10MLPs or'}a bank comprising cryopreserved MLPs.213-. (canceled)14. A method for producing platelets from megakaryocytes comprising the steps of: [ (i) TPO or a TPO agonist to cause the formation of proplatelets in culture, wherein the proplatelets release platelets; or', '(ii) hematopoietic expansion medium and optionally (1) TPO or a TPO agonist, SCF, IL-6 and IL-9 or (2) TPO or a TPO agonist, SCF, and IL-11 to cause the formation of pro-platelets in culture, wherein the pro-platelets release platelets; and, 'b) contacting the megakaryocytes with'}, 'c) isolating the platelets', 'optionally wherein the non-adherent culture of megakaryocytes is feeder-free, non-adherent culture that includes a cell population comprising megakaryocytes positive for CD41a and CD42b expression ...

Modified caspase-9 polypeptides and methods of use thereof

Provided herein are modified caspase-9 polypeptides, and chimeric caspase-9 proteins containing the modified caspase-9 polypeptides. The disclosure further provides polynucleotides encoding these proteins, engineered host cells containing these polynucleotides and proteins, including host cells that co-express a chimeric antigen receptor, and methods of making and using the same.

METHOD FOR PRODUCING ADULT LIVER PROGENITOR CELLS

Novel adult liver progenitor cells (called H2Stem Cells) have been have been characterized on the basis of a series of biological activities and markers. Methods for producing H2Stem Cells allow providing such cells in the form of adherent cells and three-dimensional cell clusters in suspension that can be differentiated into cells having strong liver-specific activities and/or that can be used for treating liver diseases or for evaluating the efficacy, the metabolism, and/or toxicity of a compound. 131-. (canceled)32. A method for evaluating the efficacy , the metabolism , the stability , and/or the toxicity of one or more exogenous components , said method comprising: [ (i) at least one hepatic marker selected from albumin, HNF-3B, HNF-4, CYP1 A2, CYP2C9, CYP2E1 and CYP3A4;', '(ii) at least one mesenchymal marker selected from Vimentin, CD90, CD73, CD44, and CD29;', '(iii) at least one liver-specific activity selected from urea secretion, bilirubin conjugation, alpha-1 -antitrypsin secretion, and CYP3A4 activity;', '(iv) Sushi domain containing protein 2 (SUSD2); and', '(v) Cytokeratin-19 (CK-19);, '(I) a cell, wherein the cell is an adult liver progenitor cell characterized in that said cell is measured positive for, '(II) an isolated cell population comprising at least 60% or between 60% and 99% or between 70% or 90% of cells of part (I);', '(III) a biological material isolated from a cell of part (I) or population of part (II), wherein the biological material is a conditioned cell culture media, a protein extract, a membrane vesicle, or any fraction thereof comprising one or more isolated proteins, nucleic acids, metabolites, and/or antigens; or', '(IV) a composition comprising a cell of part (I), a cell population of part (II), or a biological material of part (III);, '(a) providing(b) exposing said cell, said cell population, said composition, or said biological material to one or more exogenous components selected from chemical compounds, proteins, nucleic ...

PLURIPOTENT HUMAN ADIPOSE ADULT STEM CELLS: ISOLATION, CHARACTERIZATION AND CLINICAL IMPLICATIONS

Methods for the efficient isolation and use of pluripotent adipose-derived stem cells (PASCs) are provided. In certain embodiments the methods involve providing an adipose tissue sample from which the stromal vascular fraction is co-cultured with the adipocyte fraction. PASCs can be isolated with a high degree of purification without requiring an additional cell enrichment process (e.g. cell sorting). PASCs and their conditioned media can be used for tissue regeneration within hours of harvesting the adipose tissue, and without requiring cell expansion. PASCs can grow as floating individual cells, as clusters of cells, or attached to surface(s) of the culture vessel. PASCs do not produce teratomas in vivo, nor do they induce immunorejection upon transplantation, and they achieve a high efficiency in grafting. The cells and compositions can be used for cell therapy and to screen new drugs. 1. A method of ameliorating tissue damage in a subject , the method comprising administering a composition comprising pluripotent stem cells (PSCs) to the subject under conditions permitting the PSCs of the composition to divide and populate a site of tissue damage ,wherein the PSCs are isolated from adipose tissue by a method comprising:(a) releasing adipose cells, including an adipocyte fraction and a stromal vascular fraction, from an adipose tissue sample using a proteolytic enzyme that breaks the peptide bonds in collagen;(b) co-incubating the released adipocytes and the released stromal vascular fraction for 2-36 hours, wherein 4-24 hours of said co-incubation takes place under stress conditions in a medium containing a proteolytic enzyme, in the absence of nutrients, under hypoxic conditions, and decreasing the temperature to 4° c.;(c) recovering the viable cells following the co-incubation in step (b) by centrifuging to produce a cell pellet, removing supernatant containing adipocyte cell debris by aspiration, washing the cell pellet to remove the proteolytic enzyme, and ...

MODULATION OF MUSCLE AND ADIPOCYTE DISTRIBUTION AND FATE

Modulation of the ADAMTS1 signaling pathway alters the commitment of progenitor cells to a muscle fate; and increases muscle mass. 1. A method of modulating metabolism relating to insulin use , the method comprising:administering to an individual in need of modulating metabolism relating to insulin use an effective dose of a purified active fragment of human ADAMTS1 protein comprising SEQ ID NO:1, residues 258-463; thereby contacting a population of human muscle satellite cells, wherein the dose is effective to modulate metabolism relating to insulin use in the individual.2. The method of claim 1 , wherein the active fragment of human ADAMTS1 protein comprises the polypeptide of SEQ ID NO:4.3. The method of claim 2 , wherein the active fragment lacks ADAMTS1 C-terminal portion from residues 725-967.4. The method of claim 1 , wherein the active fragment is administered by intramuscular injection.5. The method of claim 1 , wherein the active fragment consists of the polypeptide of SEQ ID NO:4.6. The method of claim 1 , wherein the individual is suffering from insulin resistance claim 1 , metabolic syndrome or type II diabetes.7. The method of claim 1 , wherein the contacting is performed on an ex vivo cell population.8. The method of claim 1 , wherein the stem cell is a stem or progenitor cell with muscle potential.9. The method of claim 8 , wherein the muscle stem cell is a satellite cell.10. The method of claim 9 , wherein the satellite cell is activated by contacting with the ADAMTS1 protein or an active fragment thereof. This application claims benefit and is a Divisional of application Ser. No. 14/442,704 filed May 13, 2015, which is a 371 application and claims the benefit of PCT Application No. PCT/US2013/070389, filed Nov. 15, 2013, which claims benefit of U.S. Provisional Patent Application No. 61/726,932, filed Nov. 15, 2012, which applications are incorporated herein by reference in their entirety.This invention was made with Government support under ...

METHOD OF DOUBLE-COATING LACTIC ACID BACTERIA

The present disclosure relates to production of double-coated lactic acid bacteria using peptides and polysaccharide. Double-coated lactic acid bacteria show improved heat-resistance, acid-resistance, bile-resistance, storage stability and excellent survival rate when reaching intestine. 1. A method of preparing double-coated lactic acid bacteria , the method comprising:providing a protein aqueous solution comprising either or both of skim milk and isolated soybean protein as a protein component;adding a proteolytic enzyme to the protein aqueous solution, which causes partial hydrolysis of the protein component in the protein aqueous solution to provide a hydrolyzed product comprising water-soluble peptides and water semi-soluble peptides;cultivating lactic acid bacteria in a fermentation solution comprising the hydrolyzed product, wherein at least part of the water-soluble peptides from the hydrolyzed product are consumed by the lactic acid bacteria, wherein cultivating provides a liquid mixture comprising the lactic acid bacteria, the water-soluble peptides and the water-semisoluble peptides;centrifuging the liquid mixture to deposit the water-semisoluble peptides along with the lactic acid bacteria such that deposited water-semisoluble peptides enclose lactic acid bacteria to provide peptide-coated lactic acid bacteria,collecting, from the centrifugation result, the peptide-coated lactic acid bacteria,mixing, in an aqueous solution, the collected peptide-coated lactic acid bacteria with a polysaccharide and a cryoprotectant to provide a homogenized aqueous mixture comprising the peptide-coated lactic acid bacteria, the polysaccharide and the cryoprotectant,freezing the homogenized aqueous mixture to provide a frozen homogenized mixture in which the peptide-coated lactic acid bacteria are enclosed in a polysaccharide coating, andlyophilizing the frozen homogenized mixture, thereby forming double-coated lactic acid bacteria coated with the water-semisoluble ...

Chemical reprogramming of human glial cells into neurons with small molecule cocktail

Provided are compositions, articles and methods that relate to promoting neurogenesis or neuroregeneration in mammalian nervous system. Embodiments relate to use of groups of compounds that contain Crizotinib (Cri), Flurbiprofen, Lithium Chloride (Li), Vitamin C (VC), Ceritinib (Cer) or Pirfenidone (PFD). In certain implementations glial cells are converted into functional neurons.

A new biomarker expressed in pancreatic beta cells useful in imaging or targeting beta cells

The present invention is directed to the identification of a biomarker specifically located in the plasma membrane of pancreatic beta cells. Based on a set of specific features the biomarker is a unique candidate for imaging and targeting strategies to study the pancreatic beta cell mass in health and disease (T1D, T2D, pancreatic cancers, obesity, islet transplantation, beta cell regeneration).

Composition including Stem Cell-Derived Exosome for Inducing Adipogenic Differentiation and Adipose Tissue Regeneration

A composition for inducing differentiation into adipocytes or regenerating adipose tissues comprises, as an ingredient, exosomes derived from stem cells differentiating into adipocytes, or exosomes derived from proliferating stem cells. 1. A composition for inducing differentiation into adipocytes or regenerating adipose tissues comprising , as an ingredient , exosomes derived from stem cells differentiating into adipocytes.2. The composition of claim 1 , wherein the stem cells differentiating into the adipocyte comprise one or more of bone marrow stem cells claim 1 , cord blood stem cells claim 1 , and adipose-derived stem cells.3. The composition of claim 2 , wherein the stem cells differentiating into the adipocyte are human-derived claim 2 , animal-derived or plant-derived stem cells.4. The composition of claim 1 , wherein the exosomes are capable of inducing differentiation into adipocytes or regenerating adipose tissues from stem cells after treatment of the stem cells using the exosomes at a concentration of 1 to 150 μg per 1 mL of the composition.5. A cosmetic composition comprising the composition of .6. A medium composition for inducing differentiation into adipocytes comprising the composition of .7. An injectable preparation comprising the composition of and a hydrogel.8. A method of inducing stem cells differentiating into adipocytes claim 1 , the method comprising:preparing exosomes derived from the stem cells differentiating into adipocytes; andtreating stem cells with the exosomes for a time period.9. The method of claim 8 , further comprising:culturing the stem cells in a serum-free medium for a time period.10. The method of claim 8 , wherein the stem cells differentiating into the adipocyte comprise one or more of bone marrow stem cells claim 8 , cord blood stem cells claim 8 , and adipose-derived stem cells.11. The method of claim 10 , wherein the stem cells differentiating into the adipocyte are human-derived claim 10 , animal-derived or plant- ...

COMPOSITION FOR TREATING CHRONIC PULMONARY DISEASE, COMPRISING EXOSOME DERIVED FROM THROMBIN-TREATED STEM CELL

The present invention relates to a pharmaceutical composition for preventing or treating chronic pulmonary disease, a pharmaceutical formulation containing the same, and a method for preparing the same, the composition comprising as an active ingredient an exosome derived from thrombin-treated stem cells. The therapeutic agent is advantageous in that since the therapeutic agent is a cell-free preparation, the risk of carcinogenesis is low and there is no problem of transplant rejection reaction, and furthermore, there is no possibility of causing the occlusion of the microvascular system upon systemic administration, and since the therapeutic agent is a non-cell separating material, it is possible to develop a pharmaceutical agent as an off-the-shelf product, thereby reducing the manufacturing cost, and the therapeutic agent has an excellent therapeutic effect for chronic pulmonary disease with a low concentration of exosome by virtue of the thrombin treatment effect. 1. A method for treating a chronic pulmonary disease , comprising:administering to a subject in need thereof an effective amount of exosomes derived from thrombin-treated stem cells.2. The method according to claim 1 , wherein the stem cells are stem cells selected from the group consisting of mesenchymal stem cells claim 1 , human tissue-derived mesenchymal stromal cells claim 1 , human tissue-derived mesenchymal stem cells claim 1 , multipotent stem cells claim 1 , and amniotic epithelial cells.3. The method according to claim 2 , wherein the mesenchymal stem cells are derived from an umbilical cord claim 2 , umbilical cord blood claim 2 , bone marrow claim 2 , fat claim 2 , muscle claim 2 , nerve claim 2 , skin claim 2 , an amniotic membrane claim 2 , or a placenta.4. The method according to claim 1 , wherein the chronic pulmonary disease comprises bronchopulmonary dysplasia claim 1 , chronic bronchitis claim 1 , emphysema claim 1 , cystic fibrosis claim 1 , or peripheral small airway disease.5. The ...

THE METHOD OF AUTOLOGOUS PRIMARY HAIR FOLLICLES PREPARATION IN 3D CULTURE

The present invention relates to a method for the preparation of autologous human primary hair follicles in 3D cultures, comprising the isolation of primary fetal follicles; isolation of the patient's hair follicle cells; isolation of skin cells of the patient's scalp; extraction of growth factors from fetal follicle cells; the fibrin gel creation that contains growth factors of fetal follicles; sandwich cultivation of patient's hair follicle cells and skin of the patients scalp on or into fibrin gel that contains growth factors of fetal follicles; separation from fibrin gel the patients primary hair follicles, which can be used to treat baldness as an autologous graft. 1. A method of preparation of autologous primary hair follicles in 3D culture comprising the steps of:i. Extracting hair follicle units and scalp skin cells from patient's scalp;ii. Isolating mesenchymal stem cells from the extracted hair follicles of step (i);iii. Expanding the isolated mesenchymal stem cells from hair follicles of the patient of step (ii) in a 2D culture;iv. Separating and cyproserving scalp skin cells of patient extracted in step (i);v. Translating the multiplicated mesenchymal stem cells of the hair follicles of step (iii) and scalp skin cells of the patient of step (iv) into a 3D culture based on a fibrin gel which contain specific growth factors isolated from fetal hair follicles;vi. Lysing the fibrin gel of step (v) with plasmin after formation of primary hair follicles or centrifugation gel for hair follicles separation;vii. Purifying primary hair follicles of step (vi) and preparing the suspension of primary hair follicles for transplantation into the skin of the patient's scalp.2. The method of preparation of autologous primary hair follicles according to ; wherein extraction of fetal hair follicles growth factors is performed after medical pregnancy termination in the gestational period of 18to 20 weeks.3. The method of preparation of autologous primary hair follicles ...

IN-VITRO EMBRYONIC CULTURE MEDIUM COMPRISING AN INHIBITOR OF CATHEPSIN L AND THE USE OF THE SAME IN EMBRYO CRYOPRESERVATION

The present invention belongs to the technical field of embryo cryopreservation. In particular, the present invention relates to an in-vitro embryonic culture medium for improving the thawed recovery rate of an embryo, comprising an inhibitor of Cathepsin L inhibitor, for example, E-64d. The present invention also relates to a method for improving the freezing resistance and the thawing recovery rate of an embryo and a method for freezing an embryo. With the present invention, the freezing resistance and the thawing recovery rate of an embryo are significantly improved. 1. An in-vitro embryonic culture medium , comprising an inhibitor of Cathepsin L.2. The in-vitro embryonic culture medium according to claim 1 , wherein the inhibitor of Cathepsin L is E-64d claim 1 , E-64 claim 1 , or Z-Phe-Phe-FMK.3. The in-vitro embryonic culture medium according to claim 2 , wherein the inhibitor of Cathepsin L is E-64d claim 2 , and wherein E-64d is at a concentration of 0.5-3 μmol/L claim 2 , more preferably 0.5-2.0 μmol/L claim 2 , even more preferably 1.0-1.5 μmol/L claim 2 , and most preferably 1 μmol/L.4. The in-vitro embryonic culture medium according to claim 1 , further comprising any medium selected from the group consisting of CZB medium claim 1 , MTF medium claim 1 , KSOM medium claim 1 , CRmedium claim 1 , and CRmedium.5. The in-vitro embryonic culture medium according to claim 2 , further comprising any medium selected from the group consisting of CZB medium claim 2 , MTF medium claim 2 , KSOM medium claim 2 , CRmedium claim 2 , and CRmedium.6. The in-vitro embryonic culture medium according to claim 3 , further comprising any medium selected from the group consisting of CZB medium claim 3 , MTF medium claim 3 , KSOM medium claim 3 , CRmedium claim 3 , and CRmedium.7. A method for improving the freezing resistance and the thawing recovery rate of an embryo claim 3 , comprising incubating the embryo in an in-vitro embryonic culture medium comprising an inhibitor of ...

COMPOSITION FOR MAINTAINING PLATELET FUNCTION

Provided is a composition for maintaining a function of platelets, having as an active ingredient a pyridone derivative represented by the following general formula (I), or a salt thereof (in the formula, ring A, R, R, R, and Rrepresent the definitions given in the description). 2. The composition according to claim 1 , wherein a compound represented by the formula (I) is (+)-5-(3 claim 1 ,4-difluorophenyl)-5-[(3-methyl-2-oxopyridin-1(2H)-yl)methyl]imidazolidine-2 claim 1 ,4-dione.3. The composition according to claim 1 , wherein the composition is a reagent for maintaining a function of platelets or is an additive to a platelet-containing blood product.5. The method according to claim 4 , wherein a compound represented by the formula (I) is (+)-5-(3 claim 4 ,4-difluorophenyl)-5-[(3-methyl-2-oxopyridin-1(2H)-yl)methyl]imidazolidine-2 claim 4 ,4-dione.6. The method according to claim 4 , wherein a culture temperature in the culture system is from 35 to 39° C.8. The culture according to claim 7 , wherein a compound represented by the formula (I) is (+)-5-(3 claim 7 ,4-difluorophenyl)-5-[(3-methyl-2-oxopyridin-1(2H)-yl)methyl]imidazolidine-2 claim 7 ,4-dione.10. The blood product according to claim 9 , wherein a compound represented by the formula (I) is (+)-5-(3 claim 9 ,4-difluorophenyl)-5-[(3-methyl-2-oxopyridin-1(2H)-yl)methyl]imidazolidine-2 claim 9 ,4-dione.12. The method according to claim 11 , wherein a compound represented by the formula (I) is (+)-5-(3 claim 11 ,4-difluorophenyl)-5-[(3-methyl-2-oxopyridin-1(2H)-yl)methyl]imidazolidine-2 claim 11 ,4-dione. The present invention relates to a composition for maintaining a function of platelets, a method for preparing platelets, a platelet-containing blood product, and a method for maintaining a function of platelets in a blood product, which utilize a pyridone derivative.Platelets are essential for blood coagulation (hemostasis). Accordingly, there is an extremely high demand for platelets for leukemia, bone ...

Method for producing intestinal organoid derived from pluripotent stem cells

An object of the present invention is to prepare a functional intestinal organoid from pluripotent stem cells. An intestinal organoid is prepared from pluripotent stem cells, by the following steps (1) to (4): (1) differentiating pluripotent stem cells into endoderm-like cells; (2) differentiating the endoderm-like cells obtained in step (1) into intestinal stem cell-like cells; (3) culturing the intestinal stem cell-like cells obtained in step (2) to form spheroids; and (4) differentiating the spheroids formed in step (3) to form an intestinal organoid, the step including culture in the presence of a MEK1/2 inhibitor, a DNA methylation inhibitor, a TGF-β receptor inhibitor, and a γ-secretase inhibitor, in addition to an epidermal growth factor, a BMP inhibitor, and a Wnt signal activator.

Method of Deriving Mesenchymal Stem Cells

We describe a method of obtaining a cell culture, the method comprising providing a cell obtained by dispersing a human embryonic stem cell (hESC) colony, or a descendent thereof, and propagating the cell in the absence of a feeder cell layer in a serum free medium comprising FGF2 and optionally PDGF AB. Preferably, the human embryonic stem cell (hESC) colony is dispersed with a dispersing agent which is trypsin. 154-. (canceled)55. A cell line obtained by providing a cell obtained by dispersing an embryonic cell colony or a descendent thereof and propagating the cell in the absence of co-culture in a serum free medium comprising FGF2.56. A method of conditioning a cell culture medium , the method comprising obtaining a cell by dispersing an embryonic cell colony or a descendent thereof , propagating the cell in the absence of co-culture in a serum free medium comprising FGF2 , and culturing a mesenchymal stem cell , mesenchymal stem cell line or differentiated mesenchymal stem cell in a cell culture medium.57. A conditioned medium obtained by obtaining a cell by dispersing an embryonic cell colony or a descendent thereof , propagating the cell in the absence of co-culture in a serum free medium comprising FGF2 , and culturing a mesenchymal stem cell , mesenchymal stem cell line or differentiated mesenchymal stem cell in a cell culture medium.58. The conditioned medium according to claim 57 , wherein said medium is used to treat a disease in an individual.59. The cell line according to claim 55 , wherein said embryonic cell colony is a human embryonic cell colony.60. The cell line according to claim 55 , wherein said embryonic cell colony is dispersed with trypsin.61. The cell line according to claim 55 , having a cell surface marker selected from the group consisting of CD105 and CD73.62. A cell line obtained by providing a cell obtained by dispersing an embryonic cell colony or a descendent thereof claim 55 , propagating the cell in the absence of co-culture in a ...

AGENTS AND METHODS FOR TREATING AND PREVENTING SEBORRHEIC KERATOSIS

Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex vivo. Also provided herein are screening assays using cultured seborrheic keratosis cells and methods for treating seborrheic keratosis in a subject. 1. A method for culturing seborrheic keratosis cells ex vivo , the method comprising:(a) contacting a biological sample comprising seborrheic keratosis cells obtained from a subject with a solution comprising a dispase enzyme at a temperature and for a time sufficient to initiate dissociation of the seborrheic keratosis cells from the biological sample, and(b) culturing the dissociated seborrheic keratosis cells.2. The method of claim 1 , wherein the temperature is below a standard room temperature of 21° C.3. The method of claim 1 , wherein the time sufficient to initiate digestion of the seborrheic keratosis cells is at least 15 hours.4. The method of claim 1 , further comprising a step of contacting the biological sample comprising seborrheic keratosis cells with an additional protease.5. The method of claim 4 , wherein the additional protease is Trypsin.6. The method of claim 1 , further comprising a step of adding a culture medium and filtering larger particles from the dissociated cells before the culturing of step (b).7. The method of claim 6 , wherein the dissociated cells are cultured on coated plates. This application is a Continuation of U.S. application Ser. No. 15/254,344, filed on Sep. 1, 2016, which is a Divisional of U.S. application Ser. No. 14/395,737 (now U.S. Pat. No. 9,458,462), filed on Oct. 20, 2014, which is a 35 U.S.C. 371 National Stage Application of International Application No. PCT/US2013/038358, filed on Apr. 26, 2013, which designates the United States, and which claims benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 61/638,684, filed on Apr. 26, 2012, the contents of each of which are incorporated herein by reference in their entireties.Human skin is continuously exposed ...

COMPOSITIONS, METHODS AND USES OF ALPHA 1-ANTITRYPSIN FOR EARLY INTERVENTION IN BONE MARROW TRANSPLANTATION AND TREATMENT OF GRAFT VERSUS HOST DISEASE

Embodiments of the present invention relate to compositions and methods for treatment of subjects in need of or having a bone marrow transplant. Certain embodiments describe compositions and methods for treatment of conditions associated with bone marrow transplantations in a subject, for example, Graft versus Host Disease (GvHD) or bone marrow transplantation rejection. Some embodiments concern early or immediate bone marrow transplantation rejection. Certain embodiments relate to compositions and uses of alpha1-antitrypsin (α1-antitrypsin, AAT) and carboxyterminal peptide derivatives thereof and/or compositions and uses of serine protease inhibitors, immunomodulators or anti-inflammatory agent activity similar to that of AAT. 1. A method for treating or preventing acute graft versus host disease in a subject in need thereof , said method comprising administering to the subject a composition comprising alpha 1-antitrypsin (AAT) or mutant thereof or a carboxyterminal derivative thereof.2. The method of claim 1 , wherein the composition is administered to the subject before transplantation claim 1 , during transplantation claim 1 , after transplantation or combination thereof3. The method of claim 1 , wherein the composition further comprises one or more anti-transplant rejection agent claim 1 , anti-inflammatory agent claim 1 , immunosuppressive agent claim 1 , immunomodulatory agent claim 1 , anti-microbial agent claim 1 , or a combination thereof.4. The method of claim 1 , wherein administration occurs within 30 to 40 days after bone marrow transplantation.5. The method of claim 1 , wherein administration occurs immediately after bone marrow transplantation.6. The method of claim 1 , further comprising treating the subject soon after transplantation and preventing mortality of the subject compared to a subject not treated with the composition.7. A method for reducing or preventing bone marrow transplant rejection in a subject having or in need of a bone marrow ...

Method Of Selecting Stem Cells Having Ability To Produce Extracellular Vesicles With High Efficiency Using Activation Of Protease-Activated Receptor-Mediated Signaling Pathways

The present disclosure relates to a method of selecting stem cells having the ability to produce extracellular vesicles with high efficiency, the method including the step of measuring the activity of protease-activated receptor (PAR)-mediated signaling pathways, stem cells selected by the method, and a method of screening an inducer for the production of extracellular vesicles. According to the present disclosure, upon treatment of stem cells with thrombin, the production of extracellular vesicles in the stem cells and the levels of proteins in the extracellular vesicles are significantly increased via PAR-mediated signaling pathways, and thus stem cells having the ability to produce extracellular vesicles with high efficiency can be efficiently selected by treating stem cells with thrombin and measuring an activation level of a PAR-mediated signaling pathway, and stem cells selected by this method can be effectively used in related research and clinical fields.

METHODS FOR THE PREPARATION OF FIBROBLASTS

The invention relates to a process for generating fibroblasts, more particularly, to the culturing of fibroblasts in large numbers and of the heterogenic type. The invention is also directed to the use of fibroblasts in the preparation of heterotypic spheroids and a process for the preparation of such heterotypic spheroids. 1. A process for the preparation of fibroblasts comprising the steps of:a) Providing a cell-containing tissue sample;b) Preparing a suspension of primary cells from the cell-containing tissue sample;c) Culturing the suspension of primary cells wherein fibroblast cell nests are generated from and within the suspension of primary cells;d) Separating the fibroblast cell nests of step (c) from the suspension of primary cells;e) Repeating steps (c) to (d) at least once.2. A process according to claim 1 , wherein the suspension of primary cells of step b) is also treated with an enzymatic composition containing one or more enzymes selected from the group consisting of proteases such as serin proteases such as trypsin or dispases claim 1 , neutral proteases; metalloendopeptidases such as collagenases such as interstitial collagenases and neutrophil collagenases or thermolysin; DNases; hyaloronidases; before culturing according to step c).3. A process according to claim 2 , wherein the enzymatic composition also contains a serum-free medium selected from the group consisting of RPMI claim 2 , DMEM claim 2 , F15 claim 2 , MEM claim 2 , BMEEARL claim 2 , HAMFSF-12 claim 2 , Leibovitz L-15 claim 2 , McCoys 5A claim 2 , medium 199 claim 2 , Waymouth medium and HANK-solution.4. A process according to claim 1 , wherein the cell containing tissue sample is either benign tissue such as gastric tissue claim 1 , colorectal tissue claim 1 , liver tissue claim 1 , lung tissue claim 1 , mucosal tissue claim 1 , cerebral tissue claim 1 , pancreas tissue claim 1 , hepatic tissue claim 1 , dermal tissue claim 1 , prostate or periprostatic tissue claim 1 , gastric tissue ...

METHOD FOR RAPID MINCING OF ADIPOSE TISSUES TO ISOLATE LIVE CELLS IN VITRO

A method for rapid mincing of adipose tissues to isolate live cells in vitro is disclosed to overcome the drawbacks of a lead procedure of isolating live cells by holding a knife to mince adipose tissues, which can only isolate a small number of live cells and obtain low cell viability. The method includes: providing an adipose tissue; mincing the adipose tissue homogeneously by a mincing device; adding a reagent into the minced adipose tissue to perform hydrolysis; performing centrifuge and isolation; and removing a supernatant to obtain a cell pellet. Therefore, the time of mincing adipose tissues can be shortened, and contaminations caused by repeated use of the knife can be avoided. The method can be used for isolating live cells of adipose tissues to improve the number of live cells per unit weight of adipose tissues without reducing the cell viability. 1. A method for rapid mincing of adipose tissues to isolate live cells in vitro , comprising the steps as follows:step (a) providing an adipose tissue;step (b) arranging the adipose tissue into an enclosed vessel of a cutting device;step (c) applying an acting force on the enclosed vessel by the cutting device to enable a preset cutting tool in the enclosed vessel to implement rapid and homogeneous mincing on the adipose tissue, thereby obtaining a homogenized adipose tissue;step (d) adding a reagent into the homogenized adipose tissue to implement a hydrolysis reaction;step (e) implementing filtering and centrifugal separation on the homogenized adipose tissue treated via the hydrolysis reaction; andstep (f) removing a supernate after the centrifugal separation so as to obtain a cell pellete.2. The method for rapid mincing of adipose tissues to isolate live cells in vitro according to claim 1 , wherein a step of cleaning the adipose tissue via a phosphate buffer solution may be included before step (a).3. The method for rapid mincing of adipose tissues to isolate live cells in vitro according to claim 1 , ...

HEAT-EXPOSED PLATELET LYSATE COMPOSITIONS, AND METHODS FOR PREPARING AND USING SAME

The present disclosure provides a composition derived from a platelet concentrate, methods of making the composition, and culture medium supplemented with the composition. Preferred methods of making the composition include heat treating a platelet lysate composition. 1. A method for preparing a bioactive platelet composition , the method comprising:providing a starting platelet lysate composition, the starting platelet lysate composition having less than 20,000 ng/ml fibrinogen; andheating the starting platelet lysate composition under conditions and for a period of time effective to modify a level of at least one growth factor.2. The method of claim 1 , wherein the starting platelet lysate composition includes a first level of FGF-b claim 1 , and wherein the modified platelet lysate composition includes FGF-b at a second level of no more than 10% of the first level of FGF-b.3. The method of claim 1 , wherein the starting platelet lysate composition includes a first level of VEGF claim 1 , and wherein the modified platelet lysate composition includes VEGF at a second level of at least 65% of the first level of VEGF.4. The method of claim 1 , wherein the starting platelet lysate composition includes a first level of TIMP-1 claim 1 , and wherein the modified platelet lysate composition includes TIMP-1 at a second level of at least 40% of the first level of TIMP-1.5. The method of claim 1 , wherein the starting platelet lysate composition includes a first level of PDGF-BB claim 1 , and wherein the modified platelet lysate composition includes PDGF-BB at a second level of at least 70% of the first level of PDGF-BB.6. The method of claim 1 , wherein the starting platelet lysate composition includes a first level of BMP-2 claim 1 , and wherein the modified platelet lysate composition includes BMP-2 at a second level of at least 50% of the first level of BMP-2.7. The method of claim 1 , wherein the starting platelet lysate composition includes a first level of MMP-1 claim ...

SPECIFICATION OF FUNCTIONAL CRANIAL PLACODE DERIVATIVES FROM HUMAN PLURIPOTENT STEM CELLS

Cranial placodes are embryonic structures essential for sensory and endocrine organ development. The efficient derivation of cranial placodes from human pluripotent stem cells is disclosed where the timed removal of the BMP inhibitor Noggin, a component of the dual-SMAD inhibition strategy of neural induction, triggers placode induction at the expense of CNS fates. Further fate specification at the pre-placode stage enables the selective generation of placode-derived trigeminal ganglia capable of in vivo engraftment, mature lens fibers and anterior pituitary hormone-producing cells that upon transplantation produce hormones including, but not limited to, human growth hormone and adrenocortiocotropic hormone in vivo. Alternatively, anterior pituitary hormone-producing cells are generated in cell culture systems in vitro. 1. A method for inducing differentiation of cells , comprising:a) contacting a plurality of cells with an inhibitor of Small Mothers Against Decapentaplegic protein signaling (“SMAD inhibitor”); andb) contacting the cells with a bone morphogenetic protein (“BMP”);wherein the cells are contacted with the SMAD inhibitor and the BMP in an amount effective to induce detectable expression of SIX1 and PAX6 in the plurality of cells.2. The method of claim 1 , wherein the method comprises one or more of the following:(i) the cells are contacted with the SMAD inhibitor for up to about 11 days, and wherein the cells are contacted with the BMP for up to about 3 days;(ii) the cells are contacted with an activator of Wnt signaling (“Wnt activator”) in an amount effective to induce a detectable level of expression of SIX1 and PAX3 in the plurality of cells;(iii) the cells are contacted with an activator of sonic hedgehog signaling (“SHH activator”) in an amount effective to induce a detectable level of expression of SIX1 and PITX1 in the plurality of cells; and(iv) the cells are contacted with a BMP to induce a detectable level of expression of SIX1 and PITX3 in ...

Spinal cord injury treatment neurosphere

A neurosphere for spinal cord injury treatment in which the phosphorylation of p38 MAPK is enhanced as compared to the control, where the control is a neurosphere that has not contacted with a γ-secretase inhibitor, or neurons obtained by inducing differentiation from a neurosphere that has not contacted with a γ-secretase inhibitor, is useful for the treatment of spinal cord injuries.

FUNCTIONAL CORTICO-SPINAL-MUSCLE ASSEMBLED SPHEROIDS

Functional human cortico-spinal-muscle assembled spheroids are generated by in vitro culture. Complete cortico-spinal-muscle spheroids (hCS-hSC-hSkM) are assembled from component cultured cell systems, where each cultured cell system is designed to provide specific sets of neural and/or muscle cells, and which components are functionally integrated in the assembled spheroid. 1. A method for producing functionally integrated human cortico-spinal-muscle assembled spheroids in vitro , the method comprising:inducing in a pluripotent stem cell suspension culture a neural fate to provide a spheroid of neural progenitor cells;(i) differentiating the neural progenitor cells in a spheroid to differentiate into spinal cord spheroids (hSC), and culturing the hSC with skeletal muscle cells under integrating conditions to generate hSC-hSkM spheroids;(i) differentiating the neural progenitor cells in a spheroid to differentiate into cortical spheroids (hCS);culturing the hCS and hSC-hSkM under conditions permissive for spheroid fusion while maintaining for an extended period of time in neural medium; wherein an integrated forebrain structure is differentiated comprising interacting glutamatergic projection neurons, motor neurons, interneurons, and muscle cells.2. The method of claim 1 , wherein the neurons comprise at least one allele associated with a neurologic or neuromuscular disorder.3. The method of claim 1 , wherein the pluripotent stem cells are induced pluripotent stem cells.4. The method of claim 1 , wherein the pluripotent stem cell suspension culture is induced to a neural fate by culturing intact colonies of the pluripotent stem cells in medium comprising an effective dose of one or more SMAD inhibitor.5. The method of claim 4 , wherein the medium comprises a dose of dorsomorphin and SB-431542 effective to induce pluripotent stem cells to a neural fate.6. The method of claim 5 , wherein the suspension culture is feeder layer free.7. The method of claim 1 , wherein ...

METHOD FOR PRODUCING ADULT LIVER PROGENITOR CELLS

Novel adult liver progenitor cells (called H2Stem Cells) have been have been characterized on the basis of a series of biological activities and markers. Methods for producing H2Stem Cells allow providing such cells in the form of adherent cells and three-dimensional cell clusters in suspension that can be differentiated into cells having strong liver-specific activities and/or that can be used for treating liver diseases or for evaluating the efficacy, the metabolism, and/or toxicity of a compound. 146-. (canceled)47. An adult liver progenitor cell characterized in that said cell is measured:(a) positive for α-smooth muscle actin (ASMA), albumin (ALB), and CD140b; and(b) negative for Sushi domain containing protein 2 (SUSD2) and Cytokeratin-19 (CK-19).48. The cell of wherein said cell is further measured positive for:(a) At least one hepatic marker selected from albumin, HNF-3B, HNF-4, CYP1A2, CYP2C9, CYP2E1 and CYP3A4;(b) At least one mesenchymal marker selected from Vimentin, CD90, CD73, CD44, and CD29;(c) At least one liver-specific activity selected from urea secretion, bilirubin conjugation, alpha-1-antitrypsin secretion, and CYP3A4 activity;(d) At least one marker selected from ATP2B4, ITGA3, TFRC, SLC3A2, CD59, ITGB5, CD151, ICAM1, ANPEP, CD46, and CD81; and(e) At least one marker selected from MMP1, ITGA11, FMOD, KCND2, CCL11, ASPN, KCNK2, and HMCN1.49. The cell of characterized in that the cell is measured negative for:(a) CD271;(b) At least one marker selected from ITGAM, ITGAX, IL1 R2, CDHS, and NCAM1; and(c) At least one marker selected from HP, CP, RBP4, APOB, LBP, ORM1, CD24, CPM, and APOC1.50. The cell of wherein said cell is further measured positive for at least one liver-specific activity selected from sulfotransferase activity claim 47 , tryptophan-2 claim 47 ,3-dioxygenase activity claim 47 , liver carboxylase activity claim 47 , ammonia metabolism claim 47 , and glycogen storage.51. An isolated cell population comprising at least 60% claim 47 , ...

PROCESS OF PREPARING CHONDROCYTE CELL SUSPENSION AND ITS USE

A process for the preparation of chondrocyte cell suspension and its use in defect site of knee or ankle or shoulder or wrist or elbow or hip of subject. 1. A process of preparing chondrocyte cell suspension comprising(a) harvesting 40 to 100 mg weight of cartilage tissue from non-weight bearing area of knee of the subject;(b) mincing the tissue from (a) followed by digesting with enzyme(s) for the isolation of chondrocyte cells;(c) mixing chondrocyte cells with nutrient medium, serum and optionally growth factors;(d) optionally seeding to enable cell multiplication until P2 stage to obtain not less than 48 million cells within four weeks;(e) centrifuging, discarding the supernatant;(f) mixing with nutrient medium;(g) analyzing and characterizing the chondrocyte cell suspension;(h) filling the characterized chondrocyte cell suspension in transparent V shaped 1 ml vials; and optionally transporting to the same subject as in (a).2. A process of preparing chondrocyte cell suspension as claimed in wherein the subject is an adult human subject.3. A process of preparing chondrocyte cell suspension as claimed in wherein the enzyme is selected from trypsin-EDTA claim 1 , Collagenase and the like.4. A process of preparing chondrocyte cell suspension as claimed in wherein the nutrient medium is selected from IMDM claim 1 , EMEM claim 1 , DMEM and the like.5. A process of preparing chondrocyte cell suspension as claimed in wherein the growth factors are selected from IGF claim 1 , TGF claim 1 , FGF and the like.6. A process of preparing chondrocyte cell suspension as claimed in wherein the seeding is done in T-25 flask and/or T-75 and/or T-150 flask and the like.7. A process of preparing chondrocyte cell suspension as claimed in wherein the transportation is at 2 to 8 degree centigrade.8Mycoplasma. A process of preparing chondrocyte cell suspension as claimed in wherein the analysis performed are Appearance claim 1 , Sterility claim 1 , claim 1 , Endotoxin claim 1 , Cell ...

CARTILAGE TISSUE PRODUCING METHOD AND CARTILAGE TISSUE

The present invention aims to provide a method for producing a cartilage tissue, which enables production of a cartilage tissue having an appropriate thickness, form, and mechanical strength, and a cartilage tissue produced by the method for producing a cartilage tissue. Provided is a method for producing a cartilage tissue including a step of seeding a collagenase-treated cartilage tissue piece in the form of a block 50 to 1,000 μm on a side onto a porous substrate composed of a bioabsorbable material. 1. A method for producing a cartilage tissue comprisinga step of seeding a collagenase-treated cartilage tissue piece in the form of a block 50 to 1,000 μm on a side onto a porous substrate composed of a bioabsorbable material.2. The method for producing a cartilage tissue according to claim 1 ,wherein the cartilage tissue piece is in the form of a block 100 to 800 μm on a side.3. The method for producing a cartilage tissue according to claim 1 ,wherein the porous substrate is a nonwoven fabric having an average pore size of 20 to 50 μm.4. The method for producing a cartilage tissue according to claim 1 ,wherein the bioabsorbable material constituting the porous substrate is polyglycolide.5. The method for producing a cartilage tissue according to claim 1 ,wherein the porous substrate is fixed to a mold composed of a bioabsorbable material to combine and integrate with the mold.6. The method for producing a cartilage tissue according to claim 5 ,wherein the bioabsorbable material constituting the mold is polycaprolactone.7. A cartilage tissue comprisinga porous substrate composed of a bioabsorbable material anda collagenase-treated cartilage tissue piece in the form of a block 50 to 1,000 μm on a side seeded on the porous substrate. The present invention relates to a method for producing a cartilage tissue, which enables production of a cartilage tissue having an appropriate thickness, form, and mechanical strength, and to a cartilage tissue produced by the method ...

COMPOSITIONS AND METHODS FOR USING CELLS TO TREAT HEART TISSUE

This document relates to compositions containing cardiogenic factors, to methods to obtain cells by culturing initial cells in the presence of such factors; and methods of administering the obtained cells to heart tissue. 1. (canceled)2. A composition for use in differentiation of stem cells into cardioprogenitor cells comprising: TGFβ-1 , a BMP polypeptide , wherein the BMP polypeptide is selected from the group consisting of: BMP2 and BMP4; α-thrombin; FGF-2; IGF-1; Activin-A; Cardiotrophin; and Cardiogenol C.3. The composition of claim 2 , wherein the BMP polypeptide is BMP4.4. The composition of claim 2 , which further comprises at least one compound selected from the group consisting of: LIF; FGF-4; VEGF-A and combinations thereof.5. The composition of claim 2 , which lacks at least one compound chosen in the group consisting of: FGF-4; LIF; and VEGF-A.6. The composition of claim 2 , wherein the composition consists essentially of TGFβ-1 claim 2 , BMP4 claim 2 , α-thrombin claim 2 , FGF-2 claim 2 , IGF-1 claim 2 , Activin-A claim 2 , Cardiotrophin and Cardiogenol C.7. The composition of claim 2 , wherein when one compound is present in said composition claim 2 , it is present in an amount of between 1 and 5 ng of said TGFβ-1 per mL claim 2 , between 1 and 10 ng of said BMP4 per mL claim 2 , between 0.5 and 5 ng of said Cardiotrophin per mL claim 2 , between 0.5 and 5 units of said α-thrombin per mL claim 2 , between 50 and 500 nM of said Cardiogenol C claim 2 , between 1 and 10 ng of said FGF-2 per mL claim 2 , between 10 and 100 ng of said IGF-1 per mL claim 2 , between 1 and 50 ng of said Activin-A per mL claim 2 , between 1 and 10 units of said LIF per mL claim 2 , between 1 and 50 ng of said VEGF-A per mL.8. The composition of claim 2 , containing 2.5 ng/mL of recombinant TGFβ-1 claim 2 , 5 ng/mL of BMP4 claim 2 , 1 ng/mL of Cardiotrophin claim 2 , 100 nM of Cardiogenol C claim 2 , used in a combinatorial fashion.9. The composition of claim 8 , further ...

CELL DIFFERENTIATION MARKER AND ITS USES

Methods of using Dub3 protein, a nucleic acid molecule coding for the protein, or an inhibitor of the activity and/or of the expression of the protein for modulating cell differentiation. 112-. (canceled)13. A method for modulating cell differentiation comprising the administration to a determined cell:Dub3 protein, said protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any variant thereof having at least 43% identity with said amino acid sequence SEQ ID NO: 1, and having ubiquitin hydrolase activity ora nucleic acid molecule coding for said protein or said variant thereof, oran inhibitor of the activity, i.e. the ubiquitin hydrolase activity and/or of the expression of said protein or said variant thereof.14. The method according to claim 13 , for modulating totipotent or pluripotent cell differentiation.15. A method for inducing dedifferentiation of differentiated cells claim 13 , the cells obtained from the dedifferentiation of differentiated cells being iPS cells claim 13 , the method comprising a step of administering to a differentiated cellsDub3 protein, said protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any variant thereof having at least 43% identity with said amino acid sequence SEQ ID NO: 1, ora nucleic acid molecule coding for said protein or said variant thereof.16. The method according to for inducing dedifferentiation of differentiated cells claim 15 , wherein said cells Dub3 protein or said nucleic acid molecule coding for said protein are associated with at least an Oct family member protein and a Sox family member protein.17. The method according to claim 15 , wherein said Dub3 protein is expressed in said iPS cells at a level corresponding to at least 2 fold lower than the expression of said Dub3 protein in totipotent cell.18. A method for inducing a spontaneous differentiation of totipotent or pluripotent cells claim 15 , comprising the administration to a determined cell of an inhibitor of ...

CHEMICAL COCKTAIL FOR INDUCING SENESCENCE IN HUMAN NEURONS TO PROMOTE DISEASE MODELING AND DRUG DISCOVERY

Provided herein are methods and compositions for inducing chemical senescence in neurons and methods of using chemically induced senescent neurons for modeling neurodegenerative disease and drug discovery. The methods include contacting human neurons with a culture medium comprising an inhibitor of DNA glycosylase 1, an autophagy inhibitor, and an HIV protease inhibitor to obtain an in vitro population of senescent neurons within about 4 days. When the neurons are obtained from a patient having a neurodegenerative disease, chemically induced senescent neurons obtained by these methods recapitulate cellular and subcellular phenotypes observed in individuals with the neurodegenerative disease. 1. An in vitro method for inducing cellular senescence in human neurons , the method comprising(a) contacting human neurons in vitro with a composition comprising one or more of an inhibitor of DNA glycosylase 1, an autophagy inhibitor, or an HIV protease inhibitor in a cell culture medium; and(b) culturing the contacted neurons in the presence of the culture medium for about two to about four days, wherein a population of chemically induced senescent (CIS) neurons results.2. The method of claim 1 , wherein the agents are SBI-0206965 claim 1 , Lopinavir claim 1 , and O151.3. The method of claim 2 , wherein the composition further comprises sodium butyrate and claim 2 , optionally claim 2 , SCR-7.4. The method of claim 1 , where the CIS neurons express senescence associated biomarker β-galactosidase and exhibit decreased expression of one or more of H3k9Me3 claim 1 , Lap2β claim 1 , and HP1γ relative to control neurons.5. The method of claim 1 , wherein the neurons are human pluripotent stem cell (hPSC)-derived neurons claim 1 , primary neurons claim 1 , or induced neurons (iNs).6. The method of claim 5 , wherein the hPSC-derived neurons derived from human embryonic stem cells (ESCs) or human induced pluripotent stem cells (iPSCs).7. The method of claim 6 , wherein the human ...

MODULATION OF MUSCLE AND ADIPOCYTE DISTRIBUTION AND FATE

Modulation of the ADAMTS1 signaling pathway alters the commitment of progenitor cells to a muscle fate; and increases muscle mass. 1. A method of enhancing myogenesis , the method comprising:contacting a stem or progenitor cell with an effective dose of an agonist of ADAMTS1;wherein myogenesis is enhanced.2. The method of claim 1 , wherein the agonist is an ADAMTS1 protein or an active fragment thereof.3. The method of claim 2 , wherein the stem cell is a stem or progenitor cell with muscle potential.4. The method of claim 3 , wherein the muscle stem cell is a satellite cell.5. The method of claim 4 , wherein the satellite cell is activated by contacting with the ADAMTS1 agonist.6. The method of claim 3 , wherein the stem or progenitor cell is a mesenchymal stem cell or pre-adipocyte.7. The method of claim 3 , wherein the stem cell is induced to adopt a muscle fate.8. The method of claim 1 , wherein the contacting is performed in vivo to an individual in need of increased muscle mass.9. The method of claim 8 , wherein the individual is suffering from a myopathy claim 8 , and wherein the dose is effective to increase muscle mass or to reduce loss of muscle mass in the individual.10. The method of claim 8 , wherein the individual is in need of modulating metabolism relating to insulin use.11. The method of claim 10 , wherein the individual is suffering from insulin resistance claim 10 , metabolic syndrome or type II diabetes.12. The method of claim 1 , wherein the contacting is performed on an ex vivo cell population.13. The method of claim 12 , wherein the stem cell is a stem or progenitor cell with muscle potential.14. The method of claim 13 , wherein the muscle stem cell is a satellite cell.15. The method of claim 14 , wherein the satellite cell is activated by contacting with the ADAMTS1 agonist.16. The method of claim 15 , further comprising the step of transplanting the activated muscle stem cells to an individual in need thereof.17. The method of claim 16 , ...

METHOD FOR PREPARING AGONIST FOR IMPROVING BOAR SPERM MOTILITY

A method for preparing an agonist for improving boar sperm motility includes: (1) collecting and infiltrating a testicular tissue of a young boar 3-5 days after born; (2) washing the tissue with PBS; (3) incubating the tissue in a cell culture medium, and centrifuging the cell suspension to remove a supernatant; (4) adding hyaluronidase and collagenase IV followed by shaking, and adding the cell culture medium and centrifuging to remove a supernatant; (5) adding trypsin and deoxyribonuclease followed by shaking, and adding the cell culture medium; (6) filtering the cell suspension by a cell sieve; (7) centrifuging to remove a supernatant, and adding the cell culture medium; (8) culturing and passaging the cells; (9) culturing and storing the cell suspension. The invention has the advantages of good application effect, low cost and simple production process. 1. A method for preparing an agonist for improving boar sperm motility , comprising:(1) collecting a testicular tissue from a young boar 3 days after born, and infiltrating the testicular tissue in normal saline containing 1% of a double-antibody at 37° C. for 10-30 min;(2) washing the infiltrated testicular tissue three times with PBS containing 1% of the double-antibody;(3) incubating the testicular tissue in DMEM/F12 media at 37° C. for 30 min to fully disperse the testicular tissue; and transferring the dispersed testicular tissue to a centrifuge tube followed by centrifugation to remove a supernatant;(4) adding 1.5 mL of hyaluronidase and 1.5 mL of collagenase IV to the centrifuge tube; shaking the centrifuge tube violently at 37° C. every other 2 min; after continuously shaken 2-3 times, adding 3 mL of DMEM/F12 media; and centrifuging the centrifuge tube to remove a supernatant;(5) adding 1.5 mL of trypsin and 1.5 mL of deoxyribonuclease to the centrifuge tube; shaking the centrifuge tube violently at 37° C. every other 2 min; after continuously shaken 2-3 times, adding 3 mL of DMEM/F12 media to the ...

COMPOSITIONS AND METHODS FOR PRODUCING AND ADMINISTERING BROWN ADIPOCYTES

The present disclosure provides compositions comprising a hydrogel and a cell adhesion ligand that enhances the differentiation of mesenchymal stem cells into brown adipocytes, and/or enhances the maintenance of brown adipocytes. In some cases, a subject composition includes cells that are embedded within the hydrogel. The present disclosure also provides methods of making and using the subject compositions. 1. A composition comprising:a) a hydrogel; andb) a cell adhesion ligand embedded within the hydrogel, wherein the cell adhesion ligand enhances at least one of: (i) the differentiation of adipose derived mesenchymal stem cells (ADMSCs) into brown adipocytes; and (ii) the maintenance of brown adipocytes.2. The composition of claim 1 , wherein the cell adhesion ligand is a syndecan-1 ligand.3. The composition of claim 2 , wherein the cell adhesion ligand comprises the amino acid sequence RKRLQVQLSIRT.4. The composition of claim 1 , wherein the cell adhesion ligand is an αβintegrin ligand.5. The composition of claim 4 , wherein the cell adhesion ligand comprises the amino acid sequence KAFDITYVRLKF.6. The composition of claim 1 , wherein the cell adhesion ligand has a length of from about 10 amino acids to about 100 amino acids.7. The composition of claim 1 , wherein the hydrogel comprises hyaluronic acid.8. The composition of claim 1 , wherein the composition comprises two or more different cell adhesion ligands.9. The composition of claim 8 , wherein the two or more different cell adhesion ligands comprise a syndecan-1 ligand and an αβintegrin ligand.10. The composition of claim 9 , wherein the ratio of the syndecan-1 ligand to the αβintegrin ligand is present in the composition in a range of from 1:2 to 1:4.11. The composition of claim 1 , further comprising an enhancement compound that enhances the differentiation of ADMSCs to into brown adipocytes or enhances the maintenance of brown adipocytes.12. The composition of claim 11 , wherein the enhancement compound ...

METHOD FOR DETERMINING THE CARDIO-GENERATIVE POTENTIAL OF MAMMALIAN CELLS

This document is related to a method for determining the cardio-generative potential of mammalian cells which comprises the assessment of a CARdiac generation Potential Index (CARPI) as a function of the quantification of the expression of genes of said cells. It also relates to a method for quantitatively assessing the modification of this cardio-generative potential and the cardiogenic potential of a treatment aiming at cellular differentiation. 1. (canceled)2. A method for preparing a composition comprising cardiopoietic cells , said method comprising the steps of:a) obtaining stem cells;b) treating the stem cells with a cardiogenic preparation to produce cardiopoietic cells; i) collecting the expression level of two or more genes in the cardiopoietic cells, wherein said genes are selected from the group consisting of the following genes: Nkx2.5; Tbx5; MEF2C; GATA4; GATA6; Mesp1; FOG1; FOG2; Flk1; and homologues thereof;', 'ii) determining a CARdiac generation Potential Index (CARPI), the CARPI being a function of the quantification of the expression level of said two or more genes of said cells;, 'c) determining the cardio-generative potential of the cardiopoietic cells, byd) selecting cardiopoietic cells having an elevated CARPI compared to a reference CARPI of stem cells that have not been treated with the cardiogenic preparation; ande) formulating the selected cardiopoietic cells into the composition.3. The method of claim 2 , wherein the stem cells are obtained from an individual suffering from a heart disease or disorder.4. The method of claim 2 , wherein said expression level of said two or more genes is quantified at the level of messenger RNAs (mRNAs) claim 2 , microRNAs claim 2 , or a combination thereof.5. The method of claim 2 , wherein said expression level of said two or more genes is quantitatively measured at the level of messenger RNAs (mRNAs).6. The method of claim 2 , wherein said cardiopoietic cells contain no detectable sarcomeric proteins.7. ...

GENETICALLY MODIFIED MESENCHYMAL STEM CELLS EXPRESSING ALPHA-1 ANTITRYPSIN (AAT)

A method for treating a subject having a medical conditions associated with inflammation and/or an unwanted immune response, and said subject does not have an alpha1-antitrypsin (AAT) deficiency, wherein the method includes administering genetically modified mesenchymal stem cells to the subject, wherein said genetically modified mesenchymal stem cells include an exogenous nucleic acid including (i) an Alpha-1 antitrypsin (AAT) encoding region operably linked to (ii) a promoter or promoter/enhancer combination. 1. A method for treating a subject having a medical conditions associated with inflammation and/or an unwanted immune response , and said subject does not have an alpha1-antitrypsin (AAT) deficiency , wherein the method comprises administering genetically modified mesenchymal stem cells to the subject , wherein said genetically modified mesenchymal stem cells comprise an exogenous nucleic acid comprising (i) an Alpha-1 antitrypsin (AAT) encoding region operably linked to (ii) a promoter or promoter/enhancer combination.2. The method according to claim 1 , wherein the exogenous nucleic acid comprises a viral vector.3. The method according to claim 2 , wherein the viral vector is a retroviral vector.4. The method according to claim 1 , wherein the promoter or promoter/enhancer combination is a constitutive promoter.5. The method according to claim 4 , wherein the constitutive promoter is a short form of the human EEF1A1 eukaryotic translation elongation factor 1 alpha 1 promoter (EFS) claim 4 , a human phosphoglycerate kinase promoter (PGK) claim 4 , or a human EEF1A1 eukaryotic translation elongation factor 1 alpha 1 promoter (EF1alpha).6. The method according to claim 1 , wherein said promoter or promoter/enhancer combination is an inducible promoter.7. The method according to claim 6 , wherein the promoter is inducible upon differentiation of said cell post-administration.8. The method according to claim 6 , wherein the promoter is an inflammation-specific ...

METHODS FOR DIAGNOSING AND/OR TREATING STERILITY

The present invention concerns a polypeptide comprising the N-terminal fragment of a cystatin, and/or a cystatin expression inducer for use for the treatment of sterility and/or infertility, and a method of diagnosing and/or predicting sterility and/or infertility of a subject and/or of a couple of subjects, which method comprises measuring the level of at least one cystatin in a biological sample of a male subject and/or measuring the level of at least one cystatin in a biological sample of a female subject, and optionally measuring further the level of at least one cathepsin in a biological sample of said male of female subject. 1. A polypeptide comprising N-terminal fragment of a cystatin , and/or a cystatin expression inducer for use for the treatment of sterility and/or infertility.2. The polypeptide comprising N-terminal fragment of a cystatin and/or cystatin expression inducer for its use according to claim 1 , wherein said cystatin is cystatin 3.3. The polypeptide comprising N-terminal fragment of a cystatin and/or cystatin expression inducer for its use according to claim 1 , wherein the polypeptide comprising N-terminal fragment of a cystatin is administered in combination with at least one N-terminal fragment of another cystatin.4. The polypeptide comprising N-terminal fragment of a cystatin and/or cystatin expression inducer for its use according to claim 1 , for treating male and/or female sterility and/or infertility.5. The polypeptide comprising N-terminal fragment of a cystatin and/or cystatin expression inducer for its use according to claim 1 , for treating sterility and/or infertility of a couple of subjects.6. The polypeptide comprising N-terminal fragment of a cystatin and/or cystatin expression inducer for its use according to claim 1 , wherein said polypeptide or cystatin expression inducer is administered locally.7. A polypeptide comprising N-terminal fragment of a cystatin claim 1 , and/or cystatin expression inducer claim 1 , for use as a ...

METHODS FOR PRODUCTION OF PLATELETS FROM PLURIPOTENT STEM CELLS AND COMPOSITIONS THEREOF

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells. 1. A pharmaceutical preparation that is suitable for use in a human patient comprising at least 10platelets , wherein the preparation is substantially free of leukocytes and wherein substantially all of the platelets are functional.2. The pharmaceutical preparation of that comprises 10-10platelets claim 1 , optionally 10 claim 1 , 10 claim 1 , 10 claim 1 , 10 claim 1 , 10or 10platelets.32. The pharmaceutical preparation of any one of - wherein the platelets have one or more of the following attributes: a mean platelet volume range of 9.7-12.8 fL; a unimodal distribution of size in the preparation; and/or a lognormal platelet volume distribution wherein one standard deviation is less than 2 μm(preferably less than 1.5 μm claims 1 , 1 μmor even 0.5 μm).43. The preparation of any one of - wherein the platelets are positive for at least one of the following markers: CD41a and CD42b.54. The preparation of any one of - wherein the platelets are human platelets.65. The preparation of any one of - wherein at least 50% claims 1 , 60% claims 1 , 70% claims 1 , 80% or 90% of the platelets are functional for at least 2 claims 1 , 3 or 4 days after storage at room temperature.7. A bioreactor having weakly adherent or non-adherent megakaryocytes that produce functional platelets without feeder cells.8. A composition comprising at least 10MLPs.9. A cryopreserved composition ...

Method for serum-free culture of chondrocytes and serum-free culture medium

To provide a method for serum-free culture of human cartilage cells and a serum-free culture medium. A method for serum-free culture of cartilage cells, said method comprising: an enzymatic treatment step for treating a human cartilage cell-containing tissue with a protease; an inhibitor-treatment step for, after the enzymatic treatment step, treating the tissue with an inhibitor for the aforesaid protease; and a culture step for, after the inhibitor-treatment step, culturing the tissue in a serum-free culture medium that contains kartogenin and/or SAG, ITS, FGF2 and hydrocortisone. A serum-free culture medium for culturing cartilage cells, said serum-free culture medium containing kartogenin and/or SAG, ITS, FGF2 and hydrocortisone.

SPECIFICATION OF FUNCTIONAL CRANIAL PLACODE DERIVATIVES FROM HUMAN PLURIPOTENT STEM CELLS

Cranial placodes are embryonic structures essential for sensory and endocrine organ development. The efficient derivation of cranial placodes from human pluripotent stem cells is disclosed where the timed removal of the BMP inhibitor Noggin, a component of the dual-SMAD inhibition strategy of neural induction, triggers placode induction at the expense of CNS fates. Further fate specification at the pre-placode stage enables the selective generation of placode-derived trigeminal ganglia capable of in vivo engraftment, mature lens fibers and anterior pituitary hormone-producing cells that upon transplantation produce hormones including, but not limited to, human growth hormone and adrenocortiocotropic hormone in vivo. Alternatively, anterior pituitary hormone-producing cells are generated in cell culture systems in vitro. 1. A method for inducing differentiation of cells comprisinga) providing a plurality of cells;b) contacting the plurality of cells with an inhibitor of Small Mothers Against Decapentaplegic (SMAD) protein signaling; andc) contacting the plurality of cells with a bone morphogenetic protein (BMP) active agent;wherein the plurality of cells are contacted with the inhibitor of SMAD and the BMP active agent in an amount effective to induce detectable expression of SIX1 and PAX6 in the plurality of cells.2. The method of claim 1 , wherein the method comprises one or more of the following:(i) the plurality of cells are cultured with the SMAD inhibitor for up to about 11 days, and wherein the plurality of cells are cultured with the BMP active agent for up to about 3 days;(ii) the plurality of cells are contacted with an activator of Wnt in an amount effective to induce a detectable level of expression of SIX1 and PAX3 in the plurality of cells;(iii) the plurality of cells are contacted with sonic hedgehog and one or more FGF in an amount effective to induce a detectable level of expression of SIX1 and PITX1 in the plurality of cells; and(iv) the plurality ...

Method for Culturing Pluripotent Stem Cells

Provided is a method by which cells deviated from the undifferentiated state, which emerge in a colony during culture of stem cells having pluripotency, can be removed. In one aspect, provided is a method for culturing stem cells having pluripotency, the method including performing cell culture in the presence of a substance that can inhibit cell-cell adhesion. In another aspect, provided is a method for removing cells deviated from the undifferentiated state, the cells being cells that have emerged or may possibly emerge during culture of stem cells having pluripotency, the method including performing cell culture in the presence of a substance that can inhibit cell-cell adhesion. In still another aspect, provided is a method for maintaining the undifferentiated state of stem cells having pluripotency, the method including performing cell culture in the presence of a substance that can inhibit cell-cell adhesion.

Methods for production of platelets from pluripotent stem cells and compositions thereof

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.

AUTOMATED METHODS FOR ISOLATING REGENERATIVE CELLS FROM ADIPOSE TISSUE

A method of processing an adipose tissue to collect adipose derived regenerative cells is provided, wherein the method comprises providing a vessel comprising a fluid jet mixer, introducing the adipose tissue into the vessel, introducing a buffer solution into the vessel; washing the adipose tissue using the fluid jet mixer; introducing an enzyme solution into the vessel; initiating jet mixing into the vessel comprising the adipose tissue, the enzyme solution, and the buffer solution using the fluid jet mixer to digest the adipose tissue to form a digestion product; phase-separating the digestion product into a digested buoyant fat layer and a non-buoyant aqueous layer; and collecting the non-buoyant aqueous layer comprising the adipose derived regenerative cells. A system of processing an adipose tissue to collect adipose derived regenerative cells is also provided. 1. A method of processing an adipose tissue to collect adipose derived regenerative cells , comprising:providing a vessel comprising a centrally located fluid jet mixer comprising one or more nozzles configured to generate a high velocity fluid stream, wherein the nozzles are in an upward direction to generate an upward fluid stream and a jet mixing into the vessel;introducing the adipose tissue into the vessel;introducing a buffer solution into the vessel;washing the adipose tissue using the fluid jet mixer;introducing an enzyme solution into the vessel;initiating jet mixing into the vessel comprising the adipose tissue, the enzyme solution, and the buffer solution using the fluid jet mixer to digest the adipose tissue and form a digested buoyant fat layer and a non-buoyant aqueous layer comprising adipose derived regenerative cells;phase-separating the digested buoyant fat layer and the non-buoyant aqueous layer; andcollecting the non-buoyant aqueous layer comprising the adipose derived regenerative cells by filtering through a filter unit that allows passing of the non-buoyant aqueous layer.2. The ...

METHOD FOR PROMOTING GENERATION OF STEM CELL-DERIVED EXOSOME BY USING THROMBIN

The present invention relates to a method of promoting generation of exosomes from stem cell by using thrombin. The method according to the present invention has superior effects of promoting generation of stem cell-derived exosomes and thus exosomes can be more efficiently obtained thereby compared to conventionally known methods. In addition, the method can be useful for related research. 1. A method of promoting generation of an exosome from a stem cell , the method comprising:culturing a stem cell in a medium comprising thrombin.2. The method according to claim 1 , wherein the stem cell is an embryonic stem cell or an adult stem cell.3. The method according to claim 2 , wherein the adult stem cell comprises one or more adult stem cell types selected from the group consisting of mesenchymal stem cell claim 2 , human tissue-derived mesenchymal stromal cell claim 2 , human tissue-derived mesenchymal stem cell claim 2 , multipotent stem cell claim 2 , and amniotic epithelial cell.4. The method according to claim 3 , wherein the mesenchymal stem cells are derived from one or more tissues selected from the group consisting of umbilical umbilical cord claim 3 , umbilical cord blood claim 3 , bone marrow claim 3 , fat claim 3 , muscle claim 3 , nerve claim 3 , skin claim 3 , amnion claim 3 , and placenta.5. The method according to claim 1 , wherein the medium comprises one or more selected from the group consisting of Dulbecco's Modified Eagle's Medium (DMEM) claim 1 , Minimal Essential Medium (MEM) claim 1 , Basal Medium Eagle (BME) claim 1 , RPMI 1640 claim 1 , Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-10 (DMEM/F-10) claim 1 , Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 (DMEM/F-12) claim 1 , α-Minimal essential Medium (α-MEM) claim 1 , Glasgow's Minimal Essential Medium (G-MEM) claim 1 , Isocove's Modified Dulbecco's Medium (IMDM) claim 1 , and KnockOut DMEM.6. The method according to wherein the medium comprises the thrombin at a ...

Human Airway Stem Cells in Lung Epithelial Engineering

Methods of using human airway stem cells in lung epithelial engineering, optionally wherein the cells are contacted with a gamma secretase inhibitor, bioartificial airway organs produced thereby, and the use thereof, e.g., for transplantation. Also methods of treating a bio-artificial matrix with Tenascin-C and/or fibrillin 2. 1. A method of providing a bioartificial lung organ , the method comprising:{'sup': +', '+, 'providing a population of proliferative basal stem cells from a human donor wherein the cells are Krt5p63 cells, preferably obtained from the airway of the donor;'}optionally maintaining and expanding the cells in culture for up to five passages (preferably wherein cells were passaged at 60-100%, preferably 80%, confluency), optionally in the absence of a ROCK inhibitor;providing a (cell-free) lung tissue matrix including an airway and substantial vasculature;seeding the lung tissue matrix with the stem cells through the airway, and with endothelial cells through the vasculature; andmaintaining the matrix under conditions sufficient for the formation of a functional epithelium in the airways and functional vasculature, wherein maintaining the matrix comprises providing the lung tissue matrix with wet ventilation using a liquid media comprising a notch inhibitor, preferably a gamma secretase inhibitor, for a time sufficient for a first desired degree of organ maturation to occur to produce a wet-matured organ; and optionally maintaining a substantially constant fluid level in the organ chamber during wet ventilation.2. The method of claim 1 , in which the organ chamber comprises a chamber pressure sensor and a bi-directional drainage chamber pump each controlled by a control module that controls the bi-directional drainage pump in response to data transmitted by the chamber pressure sensor.3. The method of claim 1 , further comprising preventing a transpulmonary pressure gradient by equilibrating a pressure level in the venous line with a pressure level ...

AGENTS AND METHODS FOR TREATING AND PREVENTING SEBORRHEIC KERATOSIS

Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex vivo. Also provided herein are screening assays using cultured seborrheic keratosis cells and methods for treating seborrheic keratosis in a subject. 1. A method for inducing apoptosis in a seborrheic keratosis cell , the method comprising contacting a seborrheic keratosis cell with an effective amount of a composition that inhibits Akt signaling , thereby inducing apoptosis in the cell.2. The method of claim 1 , wherein the effective amount of the composition does not substantially affect the survival of normal keratinocytes.3. The method of claim 1 , wherein the composition comprises a small molecule claim 1 , a peptide inhibitor or an RNAi molecule.4. The method of claim 1 , wherein the composition comprises an Akt-1 and/or an Akt-2 inhibitor.5. The method of claim 1 , wherein the Akt inhibitor is an ATP-competitive Akt inhibitor.6. A method for treating a seborrheic keratosis in a subject claim 1 , the method comprising administering a composition comprising a therapeutically effective amount of an siRNA that targets Akt to a subject having seborrheic keratosis.7. The method of claim 6 , wherein the composition is applied topically or administered systemically.8. The method of claim 6 , wherein the therapeutically effective amount of the composition does not substantially affect the survival or normal keratinocytes.9. The method of claim 6 , wherein the composition further comprises a pharmaceutically acceptable carrier. This application is a Continuation of U.S. application Ser. No. 15/482,899 filed on Apr. 10, 2017, which is a continuation of U.S. application Ser. No. 15/254,344, filed on Sep. 1, 2016, which is a Divisional of U.S. application Ser. No. 14/395,737 (now U.S. Pat. No. 9,458,462), filed on Oct. 20, 2014, which is a 35 U.S.C. 371 National Stage Application of International Application No. PCT/US2013/038358, filed on Apr. 26, 2013, which designates the ...

IN VITRO METHODS OF DIFFERENTIATING STEM CELLS INTO NEURONS AND NEURONS GENERATED USING THE SAME

Methods of generating spinal cord glutamatergic interneurons (V2interneurons) from human pluripotent stem cells (hPSCs) are provided. A method of the present disclosure may include culturing a first population of hPSCs in vitro in a neural induction medium that includes: a retinoic acid signaling pathway activator; a sonic hedgehog (Shh) signaling pathway activator; and a Notch signaling pathway inhibitor, wherein the culturing results in generation of a second population of cultured cells containing CHX10+ V2interneurons. Also provided are non-human animal models that include the hPSC-derived spinal cord glutamatergic interneurons, and methods of producing the non-human animal models. 1. A method of generating spinal cord glutamatergic interneurons from a population of human pluripotent stem cells (hPSCs) , comprising culturing a first population of hPSCs in vitro in a neural induction medium comprising:a retinoic acid signaling pathway activator;a sonic hedgehog (Shh) signaling pathway activator; anda Notch signaling pathway inhibitor,{'sup': '+', 'wherein the culturing results in generation of a second population of cultured cells comprising CHX10 V2a interneurons.'}2. The method of claim 1 , wherein the retinoic acid signaling pathway activator comprises a retinoic acid receptor agonist.3. The method of claim 2 , wherein the retinoic acid receptor agonist comprises retinoic acid claim 2 , or a derivative thereof.4. The method of any one of to claim 2 , wherein the Shh signaling pathway activator comprises a Smoothened agonist.5. The method of claim 4 , wherein the Smoothened agonist is purmorphamine claim 4 , or a derivative thereof.6. The method of any one of to claim 4 , wherein the Notch signaling pathway inhibitor comprises an inhibitor of Notch receptor activation.7. The method of claim 6 , wherein the inhibitor of Notch receptor activation is a Notch receptor antagonist.8. The method of claim 6 , wherein the inhibitor of Notch receptor activation comprises ...

HUMAN PLURIPOTENT STEM CELL-DERIVED BRAIN ORGANOIDS FOR CANCER MODELING AND DRUG SCREENING

The present invention relates to substantially planar vascularized brain cancer organoid and methods of using such vascularized brain cancer organoids in anti-cancer drug discovery screen. In particular, provided herein are methods of producing and using complex, highly uniform vascularized brain cancer organoids that comprise physiologically relevant human cells and have the high degree of sample uniformity and reproducibility required for use in high-throughput screening applications. 1. A method of producing a substantially planar vascularized brain cancer organoid , comprising(a) contacting a plurality of human cells to a substantially planar hydrogel, wherein the hydrogel comprises an RGD-containing peptide and a matrix metalloproteinase (MMP)-sensitive peptide, the plurality of human cells comprising one or more of neural progenitor cells, endothelial cells, pericytes, and microglia;(b) culturing the contacted hydrogel for at least 7 days until a hydrogel comprising neurons, glia, and vasculature is obtained;(c) contacting cancer cells to the cultured hydrogel of (b); and(d) culturing the cancer cell-contacted hydrogel under culture conditions that promote cancer cell proliferation, whereby a vascularized brain cancer organoid comprising a plurality of cancer cells is obtained.2. The method of claim 1 , wherein the plurality of cancer cells form a tumor.3. The method of claim 1 , wherein the cancer cells comprise lung cancer cells claim 1 , breast cancer cells claim 1 , melanoma claim 1 , colon cancer cells claim 1 , pancreatic cancer cells claim 1 , or prostate cancer cells.4. The method of claim 1 , wherein the neural progenitor cells claim 1 , endothelial cells claim 1 , pericytes claim 1 , or microglia are derived from human pluripotent stem cells.5. The method of claim 1 , wherein the hydrogel comprises polymerized polyethylene glycol (PEG).6. The method of claim 1 , wherein the hydrogel has a shear modulus of between about 100 Pa to about 1600 Pa.7. The ...

METHOD FOR CELL CULTURE

A method for stem or progenitor cell culture. More precisely, the invention relates to a method for cell culture using one or more IαI (inter-alpha trypsin inhibitor or inter-alpha inhibitor) protein(s) or part(s) thereof as a component in a cell culture media or a coating on a cell culture surface material. Furthermore the invention relates to a cell culture media and a cell culture coating/matrix provided with one or more IαI proteins(s) or part(s) thereof. 1. A method for culturing stem or progenitor cells , comprising growing said cells on an uncoated surface in a cell culture medium comprising one or more proteins from an inter alpha trypsin inhibitor (IαI) protein family or parts thereof to promote adhesion of said cells to said uncoated surface as well as self renewal of said cells.2. A method according to claim 1 , wherein the one or more IαI proteins or parts thereof are un-complexed proteins isolated from serum or a serum fraction claim 1 , produced as a recombinant protein claim 1 , synthesized chemically claim 1 , or a combination thereof.3. A method according to claim 1 , wherein said uncoated surface is an uncoated or naked culture plastic.4. A method according to claim 1 , wherein the stem cells are pluripotent stem (PS) cells.5. A method according to claim 1 , wherein the cells are of human origin.6. A method according to claim 1 , wherein the one or more IαI proteins or parts thereof are selected from IαI or IαIH2 claim 1 ,B (H2).7. A method according to claim 1 , wherein the cell culture is whole serum free and serum component free.8. A method according to claim 1 , wherein the one or more IαI proteins or parts thereof are added to a cell culture surface as a coating agent claim 1 , and wherein said culture is devoid of said proteins or parts thereof.9. A method according to claim 1 , wherein the concentration of the one or more IαI proteins or parts thereof are 0.1 μg/mL-200 μg/mL claim 1 , particularly 2-100 μg/mL claim 1 , more particularly 10- ...

Method For Production Of Large Numbers Of Cartilage Cells With Phenotype Retention

A method for production of large numbers of cartilage cells with phenotype retention intended for treatment of articular cartilage lesions or preparing viable cartilage tissue by propagation of chondrocytes from cartilage explants, human or animal, with the retention of phenotypes of the cartilage cells, in which cells are chondrocytes retaining morphologic attributes of the same or chondrocyte progenitor cells. The method includes culturing the cells in which cultured cells are organized into mature hyaline cartilage on the surface of biologic structures such as cancellous or cortical bone lamina. The method for culturing chondrocytes to produce three dimensional cellular structures consisting of cartilage cells and cell produced extracellular cartilage matrix. 1. A method for production of large numbers of cartilage cells comprises the step of:preparing viable cartilage tissue by propagation of chondrocytes from cartilage explants, human or animal, with the retention of phenotypes of the cartilage cells.2. The method according to in which cells are chondrocytes retaining morphologic attributes of the same or chondrocyte progenitor cells.3. The method according to in which cultured cells are organized into mature hyaline cartilage on the surface of biologic structures such as cancellous or cortical bone lamina.4. The method according to for culturing chondrocytes to produce three dimensional cellular structure consisting of cartilage cells and cell produced extracellular cartilage matrix.5. The method according to whereby biologic material comprising hyaline cartilage tissue is propagated on the surface of thin cancellous bone which is undecalcified claim 1 , and wherein the said cancellous bone lamina is from 0.5 to 2.0 mm in thickness.6. The method according to whereby cancellous bone lamina claim 1 , 0.5 to 2.0 mm thick is decalcified to render it flexible.7. The method according to whereby cancellous bone lamina is either demineralized or undemineralized and is ...

USE OF CELLS DERIVED FROM FIRST TRIMESTER UMBILICAL CORD TISSUE

A method of isolating a pluripotent cell from human umbilical cord is described herein. The method involves collecting a sample of umbilical cord from fetal tissue obtained at less than 13 weeks of gestation, for example a first trimester umbilical cord. The sample is treated to obtain isolated umbilical cord cells, after which the isolated umbilical cord cells are incubated. Cells obtained in this way can be differentiated for use in treating conditions of cell damage, by supplanting the function of a damaged cell in a condition such as spinal cord injury, cardiovascular injury or heart disease. 1. A method of treating a condition of cellular damage , comprising supplanting the function of a damaged cell with a pluripotent cell from a human umbilical cord , wherein the pluripotent cell is obtained by:{'b': '13', 'collecting an umbilical cord from fetal tissue obtained at less than weeks of gestation, by collecting fetal placenta tissue by surgical aspiration; and separating the umbilical cord from the fetal placenta tissue;'} washing the umbilical cord with PBS,', 'cutting the umbilical cord, comprising an artery and two vessels, into smaller cut pieces,', 'treating the cut pieces with collagenase to obtain isolated umbilical cord cells, and', 'washing the isolated umbilical cord cells with PBS; and, 'treating the umbilical cord to obtain isolated umbilical cord cells, byincubating the isolated umbilical cord cells, by suspending the isolated umbilical cord cells in a maintenance medium comprising α-MEM, penicillin-streptomycin, and amphotericin; and{'sub': '2', 'maintaining the isolated umbilical cord cells under appropriate growth conditions of 37° C., 5% CO;'}wherein prior to cutting the umbilical cord into smaller cut pieces, the umbilical cord is from 0.5-2.0 cm in length and comprises an artery;wherein the isolated umbilical cord cells express one or more transcription factor associated with undifferentiated stem cells; and the transcription factor is OCT-4, ...

ENDOCRINE PRECURSOR CELLS, PANCREATIC HORMONE-EXPRESSING CELLS AND METHODS OF PRODUCTION

Disclosed herein are cell cultures and enriched cell populations of endocrine precursor cells, immature pancreatic hormone-expressing cells and mature pancreatic hormone-expressing cells. Also disclosed herein are methods of producing such cell cultures and cell populations. 1. An in vitro method of making a population of human PDX1-positive pancreatic endoderm cells , comprising:a) obtaining a population of human definitive endoderm cells; andb) contacting the population of human definitive endoderm cells from step (a) with a retinoid, thereby making the population of human PDX1-positive pancreatic endoderm cells.2. The method of claim 1 , comprising producing the population of human definitive endoderm cells by contacting a population of human pluripotent cells with at least one growth factor of the TGFβ superfamily.3. The method of claim 2 , further comprising removing the growth factor of the TGFβ superfamily prior to contacting the population of human definitive endoderm cells from step (a) with the retinoid.4. The method of claim 1 , further comprising contacting the population of human definitive endoderm cells from step (a) with a gamma secretase inhibitor.5. The method of claim 4 , wherein the gamma secretase inhibitor is DAPT.6. The method of claim 1 , further comprising transplanting the population of human PDX1-positive endoderm cells into a host to mature into insulin secreting cells.7. The method of claim 6 , wherein the insulin secreting cells form islet-like cell clusters.8. The method of claim 7 , wherein the islet-like cell clusters comprise singly-positive insulin cells.9. The method of claim 1 , wherein the population of human PDX1-positive pancreatic endoderm cells expresses NGN3 and does not substantially express a marker selected from the group consisting of ghrelin claim 1 , glucagon claim 1 , insulin claim 1 , and islet amyloid polypeptide (IAPP).10. The method of claim 1 , further comprising contacting the population of human PDX1-positive ...

METHOD AND COMPOSITION FOR THE TREATMENT OF MODERATE TO SEVERE KERATOCONJUNCTIVITIS SICCA

The present invention relates to topical ophthalmic compositions for treating or preventing epithelial lesions or ophthalmic disorders, including dry eye or keratoconjunctivitis sicca. 1. A method for treating an epithelial lesion of the eye or ophthalmic disorder comprising: topically administering an ophthalmic composition comprising conditioned media , or concentrate of said media , from cultured conjunctiva cells or cultured corneal cells in combination with a pharmaceutically acceptable carrier to the ocular surface or immediate vicinity of an eye of a subject in need thereof2. The method of claim 1 , wherein the cultured conjunctiva cells comprise an enriched population of conjunctiva epithelial cells or an enriched population of corneal epithelial cells.3. The method of claim 2 , wherein the conjunctiva epithelial cells or corneal epithelial cells are human cells.4. The method of claim 1 , wherein said composition is applied to the ocular surface of the eye.5. The method of claim 1 , wherein said composition is applied to a region of the eye adjacent the ocular surface.6. The method of claim 1 , wherein said composition comprises one or more human growth factors.7. The method of claim 1 , wherein said composition further comprises retinol or bicarbonate.8. The method of claim 6 , wherein said one or more growth factors are selected from EGF or TGF-β or combinations thereof.9. The method of claim 6 , wherein said one or more growth factors are selected from the group consisting of GM-CSF claim 6 , IL-15 claim 6 , IL-1a claim 6 , IL-2 claim 6 , IL-4 claim 6 , IL-5 claim 6 , IL-6 claim 6 , IL-7 claim 6 , IL-8 claim 6 , MCP-1 claim 6 , TNFα claim 6 , FGF-2 claim 6 , Flt-3 claim 6 , PDGF-AA claim 6 , TGF-β1 claim 6 , TGF-β2 claim 6 , and TGF-β3 claim 6 , or combinations thereof.10. The method of claim 1 , wherein said composition comprises about 3.0-5.0 mg/ml mucin claim 1 , about 0.3-0.6 pg/ml GM-CSF claim 1 , about 0.09-0.2 pg/ml IL15 claim 1 , about 0.17-0.3 pg ...

Pluripotent stem cell that can be isolated from body tissue

Objects of the present invention are to provide a method for directly obtaining pluripotent stem cells which do not have tumorigenic property from body tissue and the thus obtained pluripotent stem cells. The present invention relates to SSEA-3 (+) pluripotent stem cells that can be isolated from body tissue.

ENDOCRINE PRECURSOR CELLS, PANCREATIC HORMONE-EXPRESSING CELLS AND METHODS OF PRODUCTION

Disclosed herein are cell cultures and enriched cell populations of endocrine precursor cells, immature pancreatic hormone-expressing cells and mature pancreatic hormone-expressing cells. Also disclosed herein are methods of producing such cell cultures and cell populations. 1. An in vitro method of making a population of human PDX1-positive pancreatic endoderm cells that express NGN3 , comprising:a) obtaining a population of human definitive endoderm cells; andb) contacting the population of human definitive endoderm cells from step (a) with a retinoid, thereby making the population of human PDX1-positive pancreatic endoderm cells that express NGN3.2. The method of claim 1 , comprising producing the population of human definitive endoderm cells by contacting a population of human pluripotent cells with at least one growth factor of the TGFβ superfamily.3. The method of claim 2 , further comprising removing the growth factor of the TGFβ superfamily prior to contacting the population of human definitive endoderm cells from step (a) with the retinoid.4. The method of claim 1 , further comprising contacting the population of human definitive endoderm cells from step (a) with a gamma secretase inhibitor.5. The method of claim 4 , wherein the gamma secretase inhibitor is DAPT.6. The method of claim 1 , further comprising transplanting the population of human PDX1-positive endoderm cells that express NGN3 into a host to mature into insulin secreting cells.7. The method of claim 6 , wherein the insulin secreting cells form islet-like cell clusters.8. The method of claim 7 , wherein the islet-like cell clusters comprise singly-positive insulin cells.9. The method of claim 1 , wherein the population of human PDX1-positive pancreatic endoderm cells does not substantially express a marker selected from the group consisting of ghrelin claim 1 , glucagon claim 1 , insulin claim 1 , and islet amyloid polypeptide (IAPP).10. The method of claim 1 , further comprising contacting the ...

COMPOSITION FOR TREATING NEONATAL HIE

The present invention relates to a pharmaceutical composition for preventing or treating neonatal hypoxic ischemic encephalopathy (HIE), comprising thrombin-treated stem cells or exosomes derived therefrom as an active ingredient, and a method for producing the same. According to the present invention, the thrombin-treated stem cell or the exosome derived therefrom has an increased expression of growth factors, immunoregulatory factors, antioxidation factors, or regeneration factors compared to a group not treated with thrombin and also enhances a neuronal apoptosis inhibitory effect, and thus has an advantage in that the therapeutic effect thereof on neonatal hypoxic ischemic encephalopathy (HIE) is excellent. 1. A pharmaceutical composition for preventing or treating neonatal hypoxic ischemic encephalopathy (HIE) , comprising thrombin-treated stem cells or exosomes derived therefrom as an active ingredient.2. The pharmaceutical composition according to claim 1 , wherein the stem cells are selected from the group consisting of mesenchymal stem cells claim 1 , human tissue-derived mesenchymal stromal cells claim 1 , human tissue-derived mesenchymal stem cells and multipotent stem cells.3. The pharmaceutical composition according to claim 2 , wherein the mesenchymal stem cells are derived from the umbilical cord claim 2 , cord blood claim 2 , bone marrow claim 2 , fat claim 2 , muscle claim 2 , nerve claim 2 , skin claim 2 , amnion or placenta.4. The pharmaceutical composition according to claim 1 , wherein the treatment of neonatal HIE is characterized by inhibition of nerve cell death.5. The pharmaceutical composition according to claim 1 , which is administered into a cerebral ventricle or blood vessel.6. The pharmaceutical composition according to claim 1 , further comprising a supplementary component selected from the group consisting of culture media claim 1 , cytokines claim 1 , growth factors and genes.7. The pharmaceutical composition according to claim 1 , ...

METHODS FOR ISOLATING DIFFERENT TYPES OF SINGLE CELLS FROM OVARY

The invention provides a method for separating a mammal early follicle to obtain a single oocyte and a single granulocyte thereof. The method is capable of separating a mammal early follicle to obtain an active single oocyte and a corresponding granulocyte thereof. The invention further provides a kit for obtaining a single oocyte and a single granulocyte thereof from a mammal early follicle. 1. A method of obtaining a single oocyte of an early follicle of a mammal and single granulosa cell thereof , comprising the steps of:(1) blunt separating an ovarian tissue of the mammal;(2) digesting the mammalian ovarian tissue fragments in the first digestive solution at about 37° C. for about 20-60 minutes, wherein said first digestive solution contains collagenase I, collagenase II, collagenase IV or a mixture thereof;(3) passing the digestive solution mixture obtained in the step (2) through a first pore size cell strainer having a pore diameter of about 40 to 100 μm and then passing the filtrate through a second pore size cell strainer having a pore size of about 8-12 μm;(4) rinsing the precipitate in the second pore cell strainer with a culture solution, and then resuspending the precipitate with a culture solution;(5) aspirating a single early follicle in the resuspended solution;(6) after washing the obtained single early follicle, digesting it in the second digestive solution for about 3-10 minutes, the second digestive solution contains trypsin or Accutase;(7) transferring the digestive solution mixture obtained in step (6) to a culture solution, then separating and obtaining a single oocyte and single granulosa cells thereof.2. The method of claim 1 , wherein it further comprises the following step:(8) using the single oocyte and/or single granulosa cells thereof for single cell assay.3. The method of claim 1 , wherein the early follicle is a primordial follicle or a primary follicle.4. The method of claim 1 , wherein said first digestive solution comprises a ...

A METHOD FOR PRODUCING BIOLOGIC PRODUCT VARIANTS

Disclosed are methods for producing protein variants using multiple culture conditions and perfusion production. 1. A method of making a plurality of variant preparations , the plurality comprising at least a preparation of a first variant of a product (product variant 1) and a preparation of a second variant of a product (product variant 2) , comprising:providing a population of cells in a vessel configured to allow cell culture;(a-i) culturing the population of cells in culture medium under a first condition to form conditioned culture medium containing product variant 1;(a-ii) recovering product variant 1 from culture;(a-iii) optionally adding replacement medium to the conditioned culture medium;(a-iv) optionally further culturing the population of cells under the first condition to produce additional conditioned medium;(a-v) optionally recovering additional product variant 1;(a-vi) optionally combining product variant 1 from (a-ii) and (a-v);(b-i) culturing a population of cells in culture medium under a second condition to form conditioned culture medium containing product variant 2;(b-ii) recovering product variant 2 from culture;(b-iii) optionally adding replacement medium to the conditioned culture medium,(b-iv) optionally further culturing the population of cells under the second condition to produce additional conditioned medium.(b-v) optionally recovering additional product variant 2;(b-vi) optionally combining product variant 2 from (b-ii) and (b-v);obtaining product variant 1 from a batch of product variant 1;obtaining product variant 2 from a batch of product variant 2;thereby providing a plurality of variant preparations, the plurality comprising at least a preparation of a first variant of a product (product variant 1) and a preparation of a second variant of a product (product variant 2),wherein variant 1 (or a preparation of variant 1) differs from variant 2 (or a preparation of variant 2) by a physical, chemical, biological, or pharmaceutical ...

POLYPEPTIDES HAVING PROTEOLYTIC ACTIVITY AND USE THEREOF

The present invention relates to isolated polypeptides having proteolytic activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors and host cells comprising the polynucleotides, compositions comprising the polypeptides, and methods of producing and using the polypeptides. 1. An isolated or purified polypeptide having proteolytic activity , selected from the group consisting of: i. a polypeptide having at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to SEQ ID NO: 21,', 'ii. a polypeptide having at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to SEQ ID NO: 3,', 'iii. a polypeptide having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to SEQ ID NO: 6,', 'iv. a polypeptide having at least 81%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to SEQ ID NO: 9,', 'v. a polypeptide having at least 60%, at least 65%, at least 70%, at least 75, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to SEQ ID NO: 12, or', 'vi. a polypeptide having at least 55%, at least 60%, at least 65%, at least 70%, at least 75, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to SEQ ID NO: 18,, '(a)'}(b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 19, 1, 4, 7, 10, 13 or 16;(c) a polypeptide derived from a polypeptide of SEQ ID NO: 21, 3, 6, 9, 12, 15 or 18 by substitution, deletion or addition of one or several amino acids ...

Cosmetic Composition for Anti-aging or Anti-wrinkles Comprising Neural Stem Cell Conditioned Medium and Method for Preparing the Same

본 발명은 진피내 콜라겐 분해에 관여하는 MMPs (matrix metalloproteinases)의 발현 및 활성을 억제할 수 있는 TIMP-1 및 TIMP-2를 고농도로 함유하는 피부주름 개선능 또는 피부탄력 증진능이 우수한 신경줄기세포 배양액의 제조방법에 관한 것이다. 본 발명에 따른 고농도의 TIMP-1, TIMP-2 및 저농도의 다양한 MMPs를 유효성분으로 함유하는 신경줄기세포 배양액은 콜라겐 생성을 억제하는 MMPs의 발현 및 활성을 억제함으로써 콜라겐과 엘라스틴의 합성을 회복시키므로, 피부주름 개선 또는 피부탄력 증진용 조성물로 유용하다. The present invention is a neuronal stem cell culture solution excellent in skin wrinkle improvement or skin elasticity enhancement that contains a high concentration of TIMP-1 and TIMP-2 capable of inhibiting the expression and activity of MMPs (matrix metalloproteinases) involved in collagen decomposition in the dermis. It relates to a manufacturing method. The neural stem cell culture medium containing high concentrations of TIMP-1, TIMP-2 and low concentrations of various MMPs according to the present invention restores the synthesis of collagen and elastin by inhibiting the expression and activity of MMPs that inhibit collagen production, It is useful as a composition for improving skin wrinkles or enhancing skin elasticity.

用于乙肝病毒感染的原代肝细胞来源的肝前体样细胞模型、制备方法及应用

本发明属于生命科学和医学中乙型肝炎病毒感染细胞模型领域，提供了一种用于乙肝病毒感染的原代肝细胞来源的肝前体样细胞模型、制备方法及应用，该肝前体样细胞模型由三维分化后的功能肝细胞组成，该三维分化后的功能肝细胞由人原代肝细胞在体外转化培养后得到的肝前体样细胞经三维培养和肝向成熟培养后得到。通过实验验证，本发明中的肝前体样细胞模型感染HBV后，能够高表达RXRA、HNF4A、NTCP等HBV感染相关基因，可用于乙肝病毒感染研究或和HBV共培养用于制备乙肝病毒感染细胞模型。