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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 7061. Отображено 100.
02-02-2012 дата публикации

Method of using stem cells to aid in diagnosis

Номер: US20120027678A1
Автор: Albert Scheller, William C. Rader
Принадлежит: Individual

The present invention provides a method for the in vitro culture of embryonic stem cells, wherein the stem cells continue to express no antigen or antigen CD117, and mostly remain undifferentiated during culture. The present invention also relates to purified preparations of embryonic stem cells and for uses of embryonic stem cells in treating a wide variety of conditions, diseases and disorders.

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Номер записи: 1
22-03-2012 дата публикации

Methods for producing nonadherent avian cell lines

Номер: US20120070893A9
Автор: Bertrand Pain, Fabienne Guehenneux
Принадлежит: Vivalis SA

The present invention relates to a method for producing avian cell lines, comprising gradual or complete withdrawal of growth factors, serum and/or feeder layer so that the established lines are adherent or nonadherent cells capable of proliferating indefinitely in a basic culture medium. The invention also relates to the cells derived from such lines which are particularly useful for the production of substances of interest.

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Номер записи: 2
19-07-2012 дата публикации

Continuous culture apparatus with mobile vessel, allowing selection of fitter cell variants and producing a culture in a continuous manner

Номер: US20120184009A1
Автор: Eudes Francois Marie De Crecy
Принадлежит: Individual

A method and device for growing plant, animal or stem cells in a continuous manner.

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Номер записи: 3
26-07-2012 дата публикации

Differentiation of Pluripotent Stem Cells

Номер: US20120190111A1
Автор: Christine Parmenter, Janet Davis, Jiajian Liu, Pascal Ghislain André Bonnet
Принадлежит: Janssen Biotech Inc

The present invention is directed to methods to differentiate pluripotent stem cells. In particular, the present invention is directed to methods and compositions to differentiate pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage comprising culturing the pluripotent stem cells in medium comprising a sufficient amount of GDF-8 to cause the differentiation of the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage.

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Номер записи: 4
26-07-2012 дата публикации

Inbred c57bl/6 es cells with high developmental capacity

Номер: US20120192299A1
Автор: Sang Yong Kim
Принадлежит: COLD SPRING HARBOR LABORATORY

Described herein are inbred B6 ES cell lines that exhibit high developmental capacities and have a number of advantages over ES cell lines already available. First, they can be used for gene targeting and have a high percentage of germline transmission when injected into diploid host blastocysts (˜50-80%). Second, these ES cell lines can successfully be used to generate live pups by tetraploid blastocyst complementation, producing a high percentage (15-20%) of mice that are entirely inbred B6 ES cell derived. Third, these ES cells lines can be used to rapidly generate mice that are homozygous for a gene of interest. These advantages indicate that the inbred B6 ES cells provided here facilitate the rapid generation of inbred B6 mouse models in a cost-effective and efficient manner.

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Номер записи: 5
27-09-2012 дата публикации

Method of oocyte cryopreservation using antifreeze protein

Номер: US20120244616A1
Автор: Byung Chul Jee, Chang Suk Suh, Jun Woo Jo
Принадлежит: Bundang Hospital Seoul National University

Disclosed is a method for cryopreserving an oocyte by adding an antifreeze protein to a cryopreservation liquid (equilibrium solution, vitrification solution). The disclosed cryopreservation method of an oocyte minimizes damage to the oocyte, which increases the survival rate of the oocyte after freezing and thawing of the oocyte, and improves a fertilization rate, and a blastocyst development ratio of the oocyte.

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Номер записи: 6
18-10-2012 дата публикации

Induction of pluripotent cells

Номер: US20120264218A1
Автор: Sheng Ding, Tongxiang Lin
Принадлежит: Scripps Research Institute

The slow kinetics and low efficiency of reprogramming methods to generate human induced pluripotent stem cells (iPSCs) impose major limitations on their utility in biomedical applications. Here we describe a chemical approach that dramatically improves (>200 fold) the efficiency of iPSC generation from human fibroblasts, within seven days of treatment. This will provide a basis for developing safer, more efficient, non-viral methods for reprogramming human somatic cells.

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Номер записи: 7
08-11-2012 дата публикации

Non-viral delivery of transcription factors that reprogram human somatic cells into a stem cell-like state

Номер: US20120282229A1
Автор: Christian Kannemeier, Joel Sae Won Marh, John Sundsmo, Kyle Howerton
Принадлежит: Individual

Disclosed herein are cellular compositions, stable continuous cell cultures, reporter cell lines, pharmaceutical preparations, cell penetrable pluripotent stem cells transcription factors and methods related thereto, related to reprogrammed somatic cells.

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Номер записи: 8
28-02-2013 дата публикации

Method for preparing induced paraxial mesoderm progenitor (ipam) cells and their use

Номер: US20130052729A1
Автор: Jerome Chal, Olivier Pourquie
Принадлежит: Individual

An ex vivo method for preparing a population of induced paraxial mesoderm progenitor (iPAM) cells includes culturing pluripotent cells in an appropriate culture medium that includes an effective amount of an activator of the Wnt signalling pathway.

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Номер записи: 9
07-03-2013 дата публикации

Cryopreservation of umbilical cord tissue for cord tissue-derived stem cells

Номер: US20130059286A1
Автор: Hsiu-Kang Chang, Wei-Yu Lo
Принадлежит: HealthBanks Biotech Co Ltd

A method of preserving an umbilical cord is disclosed. The method comprises obtaining a segment of an umbilical cord; mincing the segment of the umbilical cord into cord tissue pieces; admixing the cord tissue pieces with a cryogenic composition comprising a cryoprotectant and a protein to form a mixture; shaking the mixture for a duration of no shorter than 20 minutes and no longer than 40 minutes; and cryopreserving the mixture.

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Номер записи: 10
14-03-2013 дата публикации

UMBILICAL CORD LINING STEM CELLS AND METHODS AND MATERIAL FOR ISOLATING AND CULTURING SAME

Номер: US20130065302A1
Автор: Gonzalez Rafael, Silva Francisco J.
Принадлежит: DAVINCI BIOSCIENCES LLC

Human umbilical cord lining stem cells that are capable of differentiating into cells of the mesodermal lineage and ectodermal lineage are described, as well as methods of isolating, expanding, culturing, and cryopreserving such cells. 1. A method for isolating umbilical cord lining stem cells (ULSCs) from an umbilical cord , said method comprisinga) obtaining the lining of an umbilical cord, wherein said lining is substantially free of blood, venous tissue, and arterial tissue; andb) culturing explants of said lining on a fibronectin-coated solid substrate in the presence of a low glucose growth medium for a period of time sufficient for said ULSCs to adhere to said fibronectin-coated solid substrate, said growth medium comprising 15% fetal bovine serum, a stabilized dipeptide of L-alanyl-L-glutamine, antibiotic, and a growth factor selected from the group consisting of basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), and epidermal growth factor (EGF).2. The method of claim 1 , said growth medium further comprising insulin claim 1 , transferrin claim 1 , selenium claim 1 , and sodium pyruvate.3. The method of claim 2 , said growth medium further comprising putrescine.4. The method of claim 3 , said growth medium comprising bFGF claim 3 , LIF claim 3 , and EGF.5. The method of claim 1 , wherein said antibiotic is gentamycin.6. The method of claim 1 , wherein said antibiotic is penicillin and streptomycin.7. The method of claim 1 , wherein the upper surface of each said explant is in contact with a solid substrate.8. The method of claim 1 , said method further comprising washing said cells adhered to said fibronectin-coated solid substrate.9. A composition for culturing ULSCs claim 1 , said composition comprising:a) a low glucose growth medium;b) 10% to 20% serum;c) 0.7 to 1.5% of a stabilized dipeptide of L-alanyl-L-glutamine;d) 1 to 100 ng/mL of a growth factor selected from the group consisting of basic fibroblast growth factor (bFGF), ...

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Номер записи: 11
21-03-2013 дата публикации

MEDIA AND METHODS FOR CELL CULTURE

Номер: US20130071927A1
Автор: Bradley Cara, Lu David, Lubitz Sandra, Schmidt Uli, Stojanov Tomas
Принадлежит: Sydney IVF Limited

The present invention provides a cell passaging medium comprising at least one agent capable of detaching from a surface a cell that is culture in vitro on said surface, and a water-soluble polymer capable of protecting the detached cell. The present invention also provides a cell culturing medium comprising one or more cell culture protectants capable of protecting cells in culture. The present invention further relates to the use of said media in methods for culturing cells in vitro or for deriving monolayer cell cultures of mammalian stem cells. 1. A cell culturing medium comprising an agent that modulates myosin II and/or a pathway modulating myosin II capable of protecting cells in culture.24-. (canceled)5. The cell culturing medium according to claim 1 , wherein the agent is blebbistatin.6. The cell culturing medium according to claim 1 , wherein the agent is capable of maintaining the cells in a monolayer culture on a surface.79-. (canceled)10. A method of continuously maintaining a culture of cells growing on a first surface claim 1 , comprising repeated steps of:(i) detaching the cells from the first surface, and{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(ii) culturing said detached cells on a second surface in the cell culturing medium of .'}11. A method of culturing cells in a monolayer on a surface claim 1 , comprising the step of culturing the cells using the cell culturing medium of .12. The method according to claim 11 , wherein the cells are cultured in a monolayer on a surface comprising a feeder layer or other surface coating.13. The method according to claim 12 , wherein the feeder layer comprises fibroblasts.14. The method according to claim 12 , wherein the other surface coating is collagen or matrigel.15. A method of passaging and culturing cells claim 12 , comprising the steps of:i. detaching the cells from a first surface; and{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'ii. culturing the detached cells on a second surface using ...

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Номер записи: 12
21-03-2013 дата публикации

Inbred c57bl/6 es cells with high developmental capacity

Номер: US20130074200A2
Автор: Sang Kim
Принадлежит: COLD SPRING HARBOR LABORATORY

Described herein are inbred B6 ES cell lines that exhibit high developmental capacities and have a number of advantages over ES cell lines already available. First, they can be used for gene targeting and have a high percentage of germline transmission when injected into diploid host blastocysts (˜50-80%). Second, these ES cell lines can successfully be used to generate live pups by tetraploid blastocyst complementation, producing a high percentage (15-20%) of mice that are entirely inbred B6 ES cell derived. Third, these ES cells lines can be used to rapidly generate mice that are homozygous for a gene of interest. These advantages indicate that the inbred B6 ES cells provided here facilitate the rapid generation of inbred B6 mouse models in a cost-effective and efficient manner.

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Номер записи: 13
28-03-2013 дата публикации

CONVERSION OF VASCULAR ENDOTHELIAL CELLS INTO MULTIPOTENT STEM-LIKE CELLS

Номер: US20130078718A1
Автор: Medici Damian, Olsen Bjorn
Принадлежит: President and Fellows of Harvard College

Disclosed herein is a method of producing multipotent cells, comprising, activating ALK2 of isolated endothelial cells in a serum starved environment, to thereby produce isolated multipotent cells. Activation can be following a threshold period of serum starvation. Activating ALK2 is by contacting the isolated endothelial cells with TGFβ-2 and/or BMP4. The isolated endothelial cells may be human, such as primary vascular, primary microvascular endothelial cells, primary human umbilical vein endothelial cells (HUVEC) or primary human cutaneous microvascular endothelial cells (HCMEC). The activation of ALK2 significantly decreases expression of VE-cadherein of the cells and/or significantly increases expression of one or more of STRO-1, FSP-1, α-SMA, N-cadherin, fibronectin (FN1), Snail (SNAI1), Slug (SNAI2), ZEB-1, SIP-1, LEF-1, Twist, CD10, CD13, CD44, CD73, CD90, CD120A, and CD124. The multipotent cells may further be used to generate other cell types such as osteoblast-like cells, chondrocyte-like cells, adipocyte-like cells, neural-like cells, and myocyte-like cells, by incubating the isolated multipotent cells in the appropriate culture conditions for a period sufficient to induce differentiation. The induced cells express TIE-2. 1. A method of producing multipotent cells from endothelial cells , comprising , activating ALK2 in the endothelial cells , in a serum starved environment , to thereby produce multipotent cells.2. The method of claim 1 , further comprising subjecting the endothelial cells to a threshold period of serum starvation prior to activating ALK2.3. The method of claim 1 , wherein activating ALK2 is by contacting the endothelial cells with an agent selected from the group consisting of TGFβ-2 or an analog claim 1 , derivative or functional fragment thereof claim 1 , BMP4 or an analog claim 1 , derivative or functional fragment thereof claim 1 , and a combination thereof.4. The method of claim 3 , wherein the agent is contacted to the endothelial ...

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Номер записи: 14
28-03-2013 дата публикации

METHOD FOR INDUCING PLURIPOTENCY IN HUMAN SOMATIC CELLS WITH PRDM14 OR NFRKB

Номер: US20130078720A1
Автор: Chia Na Yu, Feng Bo, Ng Huck Hui
Принадлежит: AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH

Methods of inducing pluripotency in human somatic cells and methods of maintaining pluripotency in human embryonic stem cells (hESCs) are provided, as well as cells and uses of employing such cells. The methods comprise culturing cells in the presence of (i) OCT4 and SOX2; (ii) at least one of KLF4 and c-MYC; and at least one of PRDM14 and NFRKB. 1. A method of inducing pluripotency in a human somatic cell , the method comprising culturing the human somatic cell in the presence of (i) OCT4 and SOX2; (ii) at least one of KLF4 and c-MYC; and (iii) at least one of PRDM14 and NFRKB.2. The method of wherein said culturing comprises contacting the human somatic cell with the OCT4 and the SOX2 claim 1 , with the at least one of KLF4 and c-MYC and with the at least one of PRDM14 and NFRKB so that the OCT4 claim 1 , the SOX2 claim 1 , the at least one of KLF4 and c-MYC and the at least one of PRDM14 and NFRKB are taken up by the human somatic cell.3. The method of wherein said culturing comprises expressing the OCT4 claim 1 , the SOX2 claim 1 , the at least one of KLF4 and c-MYC and the at least one of PRDM14 and NFRKB in the human somatic cell.4. The method of wherein each of the OCT4 claim 3 , the SOX2 claim 3 , the at least one of KLF4 and c-MYC and the at least one of PRDM14 and NFRKB are expressed from an expression vector.5. The method of wherein each of the OCT4 claim 4 , the SOX2 claim 4 , the at least one of KLF4 and c-MYC and the at least one of PRDM14 and NFRKB are expressed from a viral vector.6. The method of wherein PRDM14 is expressed in the human somatic cell together with OCT4 claim 1 , SOX2 and KLF4.7. The method of wherein c-MYC is also expressed in the human somatic cell.8. The method of wherein PRDM14 is expressed in the human somatic cell together with OCT4 claim 1 , SOX2 and c-MYC.9. The method of wherein NFRKB is expressed in the human somatic cell together with OCT4 claim 1 , SOX2 and KLF4.10. The method of wherein c-MYC is also expressed in the ...

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Номер записи: 15
04-04-2013 дата публикации

CO-CULTURING MAMMALIAN EMBRYONIC STEM CELLS WITH HUMAN FORESKIN FIBROBLASTS

Номер: US20130084563A1
Автор: Amit Michal, Itskovitz-Eldor Joseph
Принадлежит: Technion Research & Development Foundation Ltd.

A cell culture comprising human foreskin cells, the human foreskin cells being capable of maintaining stem cells in an undifferentiated state when co-cultured therewith. 1. A method of maintaining mammalian pluripotent stem cells in an undifferentiated state comprising culturing the mammalian pluripotent stem cells with a human foreskin fibroblast conditioned medium.2. The method of claim 1 , wherein said mammalian pluripotent stem cells are human pluripotent stem cells.3. The method of claim 1 , wherein said mammalian pluripotent stem cells are of embryonic origin.4. The method of claim 1 , wherein foreskin cells for said human foreskin fibroblast conditioned medium are obtained from an 8-14 day old male individual.5. The method of claim 1 , wherein said culturing is effected under culturing conditions including a growth medium which comprises serum and/or serum replacement.6. The method of claim 5 , wherein said serum and/or said serum replacement are provided at a concentration of at least 10%.7. The method of claim 5 , wherein said serum and/or said serum replacement are provided at a concentration of 15%.8. The method of claim 5 , wherein said serum is human serum.9. The method of claim 1 , wherein said human foreskin fibroblast conditioned medium being capable of maintaining said mammalian pluripotent stem cells in an undifferentiated claim 1 , proliferative state through at least 87 passages.10. The method of claim 1 , wherein said culturing is effected under culturing conditions including a growth medium which comprises a factor selected from the group consisting of a growth factor claim 1 , an anti oxidant and an amino acid.11. A cell culture comprising:(i) mammalian pluripotent stem cells, wherein said mammalian pluripotent stem cells are capable of forming teratomas containing cells of the endoderm, mesoderm and ectoderm germ layers; and(ii) human foreskin fibroblast conditioned medium capable of maintaining said mammalian pluripotent stem cells in an ...

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Номер записи: 16
11-04-2013 дата публикации

Method for selecting human induced pluripotent stem cells

Номер: US20130089870A1
Автор: Kazutoshi Takahashi, Mari Ohnuki, Shinya Yamanaka
Принадлежит: KYOTO UNIVERSITY

The present invention provides a method for selecting human induced pluripotent stem (iPS) cells which can be safely used for transplantation. That is, the present invention provides a method for selecting human iPS cells having reduced differentiation resistance, comprising the steps of: (1) inducing differentiation of human iPS cells; (2) detecting remaining undifferentiated cells after the step (1); and (3) selecting human iPS cells whose rate of remaining undifferentiated cells detected in step (2) is equivalent to or not more than that of control cells.

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Номер записи: 17
18-04-2013 дата публикации

IMMORTALIZED AVIAN CELL LINES

Номер: US20130095557A1
Автор: ERBS Philippe, Kapfer Marina, Silvestre Nathalie
Принадлежит: TRANSGENE S.A.

This invention relates to immortalized avian cells, including those deposited under accession numbers 09070701, 09070702, and 09070703 at the ECACC, and to the use of these cells for the production of viruses. The cells according to the invention are particularly useful for the production of recombinant viral vectors which can be used for the preparation of therapeutic and/or prophylactic compositions for the treatment of animals and more particularly humans. 1. An immortalized avian cell line as deposited at the European Collection of Cell Cultures (ECACC) under accession number 09070701 and derivatives thereof.2. An immortalized avian cell line as deposited at the European Collection of Cell Cultures (ECACC) under accession number 09070702 and derivatives thereof.3. An immortalized avian cell line as deposited at the European Collection of Cell Cultures (ECACC) under accession number 09070703 and derivatives thereof.4. The immortalized avian cell according to claim 1 , wherein it further comprises one or more nucleic acid sequence allowing the propagation of a defective virus.5. The immortalized avian cell line according to claim 1 , wherein it further comprises a nucleic acid sequence coding a substance of interest.6. A method for replicating a virus claim 1 , wherein said method comprises employing an immortalized avian cell line according to any one of to .7. The method according to claim 6 , wherein said virus is a live virus claim 6 , an attenuated virus claim 6 , or a recombinant virus.8. The method according to claim 6 , wherein said virus is chosen from the group consisting of poxvirus claim 6 , adenovirus claim 6 , retrovirus claim 6 , herpervirus claim 6 , alphavirus claim 6 , foamy virus claim 6 , adenovirus-associated virus claim 6 , flavivirus) and influenza virus.9. The method according to claim 8 , wherein said virus is a poxvirus.10. The method according to claim 9 , wherein said virus is a vaccinia virus.11. The method according to claim 10 , ...

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Номер записи: 18
25-04-2013 дата публикации

METHODS AND COMPOSITIONS FOR GROWTH OF CELLS AND EMBRYONIC TISSUE ON A SYNTHETIC POLYMER MATRIX

Номер: US20130102023A1
Автор: Eyster Thomas, Lahann Joerg, Nandivada Himabindu, Smith Gary D., Villa Diaz Luis
Принадлежит: THE REGENTS OF THE UNIVERSITY OF MICHIGAN

The present invention provides methods and compositions for establishing and maintaining growth of cells and embryonic tissue on a synthetic polymer matrix. For example, the present invention provides synthetic growth matrices for stem cells, gametes, mature differentiated cells, and embryonic tissue (e.g., blastomeres, embryos, and embryoid bodies). In certain embodiments, the cells are capable of going through multiple passages while remaining in an undifferentiated state as a result of the synthetic polymer matrix. 1. A composition for growth and maintenance of cells or embryonic tissue comprising: a synthetic polymer matrix and a culture medium , wherein said synthetic polymer matrix is selected from the group consisting of:i) a first polymer which comprises a zwitterionic group;ii) a co-polymer comprising poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide] (PMEDSAH) and poly(ethylene glycol methacrylate) (PPEGMA);iii) a copolymer comprising PMEDSAH and poly(2-(methacryloyloxy)ethyl)trimethylammonium chloride (PMETAC); andiv) a copolymer comprising 2-hydroxyethyl methacrylate (HEMA) and PMEDSAH.4. The composition of claim 1 , wherein said first polymer comprises either zwitterionic group is poly[2-(methacryloyloxy)ethyl dimethyl(3-sulfopropyl)ammonium] (PMEDSA) or Poly[[3-(methacryloylamino)propyl]dimethyl(3-sulfopropyl)ammonium hydroxide] (PMAPDSAH).5. The composition of claim 1 , wherein said cells are stem cells claim 1 , and wherein said stem cells remain pluripotent and maintain native karyotype after at least 5 passages.6. The method of claim 5 , wherein said stem cells are selected from the group consisting of adult stem cells and embryonic stem cells.7. A method for culturing cells or embryonic tissue comprising i) a first polymer which comprises a zwitterionic group;', 'ii) a co-polymer of poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide] (PMEDSAH) and poly(ethylene glycol methacrylate) (PPEGMA);', 'iii) a ...

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Номер записи: 19
25-04-2013 дата публикации

METHODS FOR REPROGRAMMING SOMATIC CELLS

Номер: US20130102074A1
Автор: Hochedlinger Konrad, Jaenisch Rudolf
Принадлежит: WHITEHEAD INSTITUTE FOR BIOMEDICAL RESEARCH

The invention provides methods for reprogramming somatic cells to generate multipotent or pluripotent cells. Such methods are useful for a variety of purposes, including treating or preventing a medical condition in an individual. The invention further provides methods for identifying an agent that reprograms somatic cells to a less differentiated state. 1. A composition comprising an isolated primary somatic cell that comprises an exogenously introduced Oct4 protein.2. The composition of claim 1 , wherein the isolated somatic cell further comprises an exogenously introduced Sox2 or Nanog protein.3. The composition of claim 1 , wherein the isolated somatic cell is a mammalian cell.4. The composition of claim 1 , wherein the isolated somatic cell is a human cell or a mouse cell.5. The composition of claim 1 , wherein the isolated somatic cell is an adult stem cell.6. The composition of claim 5 , wherein the adult stem cell is selected from the group consisting of: a hematopoietic stem cell claim 5 , a neural stem cell claim 5 , and a mesenchymal stem cell.7. The composition of 1 claim 5 , comprising a candidate agent of interest with respect to its potential to reprogram a somatic cell.8. The composition of claim 7 , wherein the candidate agent of interest is a DNA methylation inhibitor claim 7 , a histone deacetylase inhibitor or PD098059.9. The composition of claim 7 , wherein the candidate agent of interest is an exogenously introduced nucleic acid that encodes a pluripotency protein selected from the group consisting of: Oct4 claim 7 , Nanog claim 7 , and Sox-2.10. A somatic cell reprogramming composition comprising an isolated Oct4 protein and a candidate agent of interest with respect to its potential to reprogram a somatic cell.11. The composition of claim 10 , wherein the somatic cell is a mammalian cell.12. The composition of claim 10 , wherein the somatic cell is a human cell or a mouse cell.13. The composition of claim 10 , wherein the somatic cell is an ...

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Номер записи: 20
25-04-2013 дата публикации

METHODS OF ENHANCING PLURIPOTENTCY

Номер: US20130102479A1
Автор: Han Jianyong, Lim Bing, Tam Wai-Leong
Принадлежит: AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH

We provide for the use of Tbx3 (GenBank Accession Number: NM_005996.3 (SEQ ID NO. 1), NP_005987.3 (SEQ ID NO. 2), NM_016569.3 (SEQ ID NO. 3), NP_057653.3 (SEQ ID NO. 4)) in a method of enhancing or inducing pluripotency in a cell such as a somatic cell. We describe a method of reprogramming a cell, the method comprising modulating the expression and/or activity of Tbx3 in the cell. The cell may become a pluripotent cell such as a stem cell. We further describe a method of causing a cell such as a somatic cell to display one or more characteristics of a pluripotent cell, the method comprising modulating the expression and/or activity of Tbx3 in the cell. The method may further comprise modulating the expression and/or activity of one or more, a combination of or all of Oct4, Sox2 and Klf4 in the cell. 118-. (canceled)19. A method of inducing or enhancing pluripotency in a cell , the method comprising modulating expression and/or activity of Tbx3 (GenBank Accession Number: NM005996.3 (SEQ ID NO.: 1) , NP005987.3 (SEQ ID NO.: 2) , NM016569.3 (SEQ ID NO.: 3) , NP057653.3 (SEQ ID NO.: 4)) in the cell.20. The method of claim 19 , wherein the cell is a somatic cell.21. A method of reprogramming a cell claim 19 , the method comprising modulating expression and/or activity of Tbx3 (GenBank Accession Number: NM005996.3 (SEQ ID NO.: 1) claim 19 , NP005987.3 (SEQ ID NO.: 2) claim 19 , NM016569.3 (SEQ ID NO.: 3) claim 19 , NP057653.3 (SEQ ID NO.: 4)) in the cell.22. The method of claim 21 , wherein the step of modulating comprises upregulating the expression and/or activity of Tbx3.23. The method of claim 21 , wherein the cell becomes pluripotent.24. The method of claim 23 , wherein the cell becomes a stem cell.25. A method of causing a cell to display one or more characteristics of a pluripotent cell claim 23 , the method comprising modulating the expression and/or activity of Tbx3 (GenBank Accession Number: NM005996.3 (SEQ ID NO.: 1) claim 23 , NP005987.3 (SEQ ID NO.: 2) claim ...

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Номер записи: 21
25-04-2013 дата публикации

Cytoplasmic transfer to de-differentiate recipient cells

Номер: US20130104253A1
Автор: Karen B. Chapman
Принадлежит: Individual

Methods for de-differentiating or altering the life-span of desired “recipient” cells, e.g., human somatic cells, by the introduction of cytoplasm from a more primitive, less differentiated cell type, e.g., oocyte or blastomere are provided. These methods can be used to produce embryonic stem cells and to increase the efficiency of gene therapy by allowing for desired cells to be subjected to multiple genetic modifications without becoming senescent. Such cytoplasm may be fractionated and/or subjected to subtractive hybridization and the active materials (sufficient for de-differentiation) identified and produced by recombinant methods.

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Номер записи: 22
02-05-2013 дата публикации

ISOLATED PRIMATE EMBRYONIC CELLS DERIVED FROM EXTENDED BLASTOCYSTS

Номер: US20130108597A1
Автор: Amit Michal, Itskovitz-Eldor Joseph
Принадлежит: Technion Research & Development Foundation Ltd.

An isolated primate embryonic cell is provided as well as cell cultures and cell lines derived therefrom. Also provided are methods of generating and using such cells. 1. A method of generating a lineage specific cell comprising inducing a lineage-specific cell differentiation in an isolated primate embryonic cell , wherein said isolated primate embryonic cell line exhibits for at least 20 passages an undifferentiated proliferative state , the ability to differentiate to derivatives of each of an endoderm , mesoderm , and ectoderm tissue and a double staining expression of brachyury and Octamer binding transcription factor 4 (OCT-4) , but not of SSEA-1 , thereby generating the lineage specific cell.2. The method of claim 1 , wherein said isolated primate embryonic cell line further expresses at least one cartilage marker.3. The method of claim 2 , wherein said at least one cartilage marker is selected from the group consisting of COMP claim 2 , aggrecan and collagen type II.4. The method of claim 1 , wherein said isolated primate embryonic cell line maintains a stable normal karyotype for at least one year.5. The method of claim 1 , wherein said isolated primate embryonic cell line expresses SSEA4 and TRA-1-60 markers.6. The method of claim 1 , wherein said isolated primate embryonic cell line expresses less TRA-1-81 marker than an embryonic stem cell of the same primate species not expressing brachyury using identical assay conditions.7. The method of claim 1 , wherein said isolated primate embryonic cell line is capable of colony organization of columnar epithelium with villi throughout the upper side of said colony.8. The method of claim 1 , wherein said isolated primate embryonic cell line exhibits an OCT4 protein level lower than the OCT4 protein level in an embryonic stem cell of the same primate species not expressing brachyury using identical assay conditions.9. The method of claim 1 , wherein said isolated primate embryonic cell line expresses more ...

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Номер записи: 23
09-05-2013 дата публикации

Methods of Screening Embryonic Progenitor Cell Lines

Номер: US20130115673A1
Автор: Chapman Karen, Sternberg Hal, West Michael
Принадлежит: BIOTIME, INC.

Aspects of the present invention include methods and compositions related to the production and use of numerous clonal lineages of embryonic progenitor cell lines derived from differentiating cultures of primordial stem cells with diverse molecular markers and having been cultured for >21 doublings of clonal expansion. The robustness of these clonally-purified lines, their ability to expand for >40 passages while maintaining their pattern of gene expression, lack of tumorigenicity, and their embryonic pattern of gene expression offers novel compositions and methods for modeling numerous differentiation pathways for the first time in vitro, and for the manufacture of purified product not existing in such a purified state in nature or using other manufacturing modalities. Representative progenitor cell lines described herein are capable of development into cutaneous adipocytes, blood-brain barrier cells, neuronal cells, or smooth muscle cells each with therapeutic potential. 1. An isolated clonal progenitor cell line expressing EYA4 , wherein the clonal progenitor cell line is an embryonic cutaneous progenitor cell.2. The isolated clonal progenitor cell line of claim 1 , wherein the cell line also expresses ADH1A claim 1 , ADH1B claim 1 , FABP4 claim 1 , CD36 claim 1 , PPARG claim 1 , ANGPT2 claim 1 , EBF2 AND DBC1.3. The isolated clonal progenitor cell line of claim 1 , wherein the cell line is encapsulated in a biomaterial.4. The isolated clonal progenitor cell line of claim 3 , wherein the biomaterial comprises a hydrogel.5. The isolated clonal progenitor cell line of claim 3 , wherein the biomaterial comprises hyaluronic acid.6. The isolated clonal progenitor cell line of claim 3 , wherein the biomaterial is chosen from calcium alginate claim 3 , agarose claim 3 , polylactic acid/poly-glycolic acid derivatives claim 3 , and fibrin.76. A kit comprising the cell line of -.8. A method of differentiating a clonal progenitor cell line into cutaneous adipocytes ...

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Номер записи: 24
09-05-2013 дата публикации

SCALABLE PRIMATE PLURIPOTENT STEM CELL AGGREGATE SUSPENSION CULTURE AND DIFFERENTIATION THEREOF

Номер: US20130115695A1
Автор: Schulz Thomas C.
Принадлежит: VIACYTE, INC.

The present invention relates to methods for production of undifferentiated or differentiated embryonic stem cell aggregate suspension cultures from undifferentiated or differentiated embryonic stem cell single cell suspensions and methods of differentiation thereof. 1. A roller bottle comprising a culture of primate pluripotent stem cell (pPSC) aggregates suspended in a physiologically acceptable medium.2. The roller bottle of claim 1 , wherein the pPSC aggregates have a diameter of about 100 to 300 microns3. The roller bottle of claim 1 , wherein the culture is substantially free of pPSC agglomerations having a diameter of more than 300 microns.4. The roller bottle of claim 1 , wherein the pPSC aggregates express at least one marker selected from the group consisting of OCT4 claim 1 , NANOG claim 1 , SSEA-3 claim 1 , SSEA-4 claim 1 , Tra-1-80 and Tra-1-60.5. The roller bottle of claim 1 , wherein the pPSC aggregates are human cells.6. The roller bottle of claim 5 , wherein the pPSC aggregates are selected from the group consisting of human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC).7. The roller bottle of claim 1 , wherein the primate pluripotent stem cell-derived cell aggregates in suspension are generated by a method comprising:a. providing a culture of undifferentiated pPSCs;b. seeding a roller bottle with the undifferentiated pPSCs in a culture medium that supports undifferentiated growth of the pPSCs; andc. agitating the pPSCs at low rotation speed to form pPSC aggregates, thereby generating a roller bottle comprising a culture of pPSC aggregates.8. The roller bottle of claim 7 , wherein the pPSC aggregates have a diameter of about 100 to 300 microns.9. The roller bottle of claim 7 , wherein the culture is substantially free of pPSC agglomerations having a diameter of more than 300 microns.10. The roller bottle of claim 7 , wherein the pPSC aggregates express at least one marker selected from the group consisting of OCT4 claim 7 , ...

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Номер записи: 25
16-05-2013 дата публикации

NATIVE WHARTON'S JELLY STEM CELLS AND THEIR PURIFICATION

Номер: US20130121972A1
Автор: Taghizadeh Rouzbeh R.
Принадлежит:

Noncultured Wharton's Jelly stem cells and methods of their purification, storage and use are provided. 1. A method of purifying noncultured Wharton's Jelly stem cells , the method comprising separating noncultured Wharton's Jelly stem cells from a digested tissue comprising Wharton's Jelly.2. The method of claim 1 , further comprising digesting a tissue comprising Wharton's Jelly.35-. (canceled)6. A method according to claim 2 , wherein the tissue is digested by mechanically increasing the surface area of the tissue.7. The method of claim 6 , comprising cutting the tissue.8. The method of claim 6 , comprising mincing the tissue.9. (canceled)10. A method according to claim 2 , further comprising separating digested and undigested tissue.11. The method of claim 10 , comprising sedimenting the undigested tissue.12. The method of claim 11 , wherein centrifugation is used for sedimenting the undigested tissue.13. The method of claim 10 , comprising filtering the digested tissue.14. The method of claim 13 , wherein the noncultured Wharton's Jelly stem cells are separated from the filtrate.15. A method according to claim 10 , further comprising washing or diluting the digested tissue before separating it from the undigested tissue.16. A method of purifying cultured and noncultured Wharton's Jelly stem cells claim 10 , the method comprising:{'claim-ref': {'@idref': 'CLM-00010', 'claim 10'}, "purifying noncultured Wharton's Jelly stem cells according to ; and"}culturing mesenchymal stem cells from the undigested tissue.17. The method of claim 16 , wherein the mesenchymal stem cells are cultured in digested Wharton's Jelly.18. Purified claim 16 , noncultured Wharton's Jelly stem cells substantially free of semi-solid Wharton's Jelly.19. Purified claim 16 , noncultured Wharton's Jelly stem cells at a temperature below −20° C.20. Wharton's Jelly stem cells according to claim 18 , wherein the cells have reduced CD73 and reduced CD105 levels compared to cultured mesenchymal stem ...

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Номер записи: 26
16-05-2013 дата публикации

Compositions Comprising Human Embryonic Stem Cells and Their Derivatives, Methods of Use, and Methods of Preparation

Номер: US20130122486A1
Автор: SHROFF Geeta
Принадлежит:

The present invention relates to a pharmaceutical composition comprising of preparations of human embryonic stem (hES) cells and their derivatives and methods for their transplantation into the human body, wherein transplantation results in the clinical reversal of symptoms, cure, stabilization or arrest of degeneration of a wide variety of presently incurable and terminal medical conditions, diseases and disorders. The invention further relates to novel processes of preparing novel stem cell lines which are free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor and conditioned media. This invention further relates to the isolation, culture, maintenance, expansion, differentiation, storage, and preservation of such stem cells. 142.-. (canceled)43. A method of partially differentiating hES cells , derivatives of hES cells , or combinations thereof , free of feeder cells , animal products or growth factors other than a βhCG agonist or a progestin , leukaemia inhibitory factor , supplementary mineral combinations , amino acid supplements , vitamin supplements , fibroblast growth factor , membrane associated steel factor , soluble steel factor , and conditioned media , comprising:(a) introducing hES cells, derivatives of hES cells, or combinations thereof in a cell culture medium consisting of minimal essential medium; and(b) incubating the cells at a temperature of about 34° C. to about 38° C. in an environment of about 3.5% to about 6% carbon dioxide for about 12 hours to about 48 hours.44. The method of claim 43 , wherein said cell culture medium is RPMI or DMEM.45. The method of claim 43 , wherein said incubation is carried out in a water jacketed cell culture incubator.46. The method of claim 43 , wherein said cells in culture medium are incubated in a biocompatible container ...

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Номер записи: 27
16-05-2013 дата публикации

METHOD FOR INDUCING DIFFERENTIATION OF HUMAN PLURIPOTENT STEM CELL INTO INTERMEDIATE MESODERM CELL

Номер: US20130122536A1
Автор: Kajiwara Masatoshi, Mae Shin-ichi, Osafune Kenji
Принадлежит: KYOTO UNIVERSITY

The present invention relates to: a method for producing an intermediate mesoderm cell from a human pluripotent stem cell, comprising a step of culturing the human pluripotent stem cell in a medium containing Activin A and Wnt or a functional equivalent of Wnt and a step of culturing cells in a medium containing BMP and Wnt or a functional equivalent of Wnt; to a method for producing a metanephric cell from the intermediate mesoderm cell produced by the first method; to a human pluripotent stem cell having a foreign reporter gene in the chromosome wherein the gene is expressed interlocked with the expression of endogenous OSR1; to a method for screening for an inducer for differentiation into intermediate mesoderm using the human pluripotent stem cell; and to a kit for inducing the differentiation into an intermediate mesoderm cell. 1. A method for producing an intermediate mesoderm cell from a human pluripotent stem cell , comprising the following steps (i) and (ii) of:(i) culturing the human pluripotent stem cell in a medium containing Activin A and Wnt or a functional equivalent of Wnt, and then(ii) culturing the cell obtained in the step (i) in a medium containing BMP and Wnt or a functional equivalent of Wnt.2. The method of claim 1 , wherein the intermediate mesoderm cell is an OSR1-positive cell.3. The method of claim 1 , wherein in the culture of the step (i) claim 1 , the human pluripotent stem cell is cultured in suspension to form a cell population or cell mass of the human pluripotent stem cell claim 1 , and in the culture of the step (ii) claim 1 , the cell population or the cell mass is subjected to adhesion culture claim 1 , thereby forming the intermediate mesoderm cell.4. The method of claim 1 , further comprising claim 1 , in the culture of the step (i) claim 1 , substantially separating the cell population or cell mass of human pluripotent stem cell into respective cells.5. The method of claim 4 , wherein step (i) further comprises adhering the ...

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Номер записи: 28
16-05-2013 дата публикации

Compositions Comprising Human Embryonic Stem Cells and Their Derivatives, Methods of Use, and Methods of Preparation

Номер: US20130122588A1
Автор: SHROFF Geeta
Принадлежит:

The present invention relates to a pharmaceutical composition comprising of preparations of human embryonic stem (hES) cells and their derivatives and methods for their transplantation into the human body, wherein transplantation results in the clinical reversal of symptoms, cure, stabilization or arrest of degeneration of a wide variety of presently incurable and terminal medical conditions, diseases and disorders. The invention further relates to novel processes of preparing novel stem cell lines which are free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor and conditioned media. This invention further relates to the isolation, culture, maintenance, expansion, differentiation, storage, and preservation of such stem cells. 130.-. (canceled)31. A method of expanding hES cells , derivatives of hES cells , or combinations thereof , free of feeder cells , animal products or growth factors other than a βhCG agonist or a progestin , leukaemia inhibitory factor , supplementary mineral combinations , amino acid supplements , vitamin supplements , fibroblast growth factor , membrane associated steel factor , soluble steel factor , and conditioned media , comprising:(a) introducing hES cells, derivatives of hES cells, or combinations thereof in a cell culture medium consisting of minimal essential medium, a progestin, and a β-human chorionic gonadotropin (βhCG) agonist; and(b) incubating the cells at a temperature of about 34° C. to about 38° C. in an environment of about 3.5% to about 6% carbon dioxide for about 12 hours to about 48 hours.32. The method of claim 31 , wherein said cell culture medium is RPMI.33. The method of claim 31 , further comprising:(c) taking an aliquot of incubated cells of (b) wherein said aliquot contains at least one cell,(d) resuspending the cells in cell ...

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Номер записи: 29
06-06-2013 дата публикации

METHODS OF GROWING AN EMBRYO TO A BLASTOCYTE STAGE OF DEVELOPMENT

Номер: US20130143317A1
Автор: Assou Said, Hamamah Samir
Принадлежит:

The present invention relates generally to the fields of reproductive medicine. More specifically, the present invention relates to a novel human embryo co-culture system to improve human embryo growth in vitro and, consequently, increase pregnancy rates in infertile women undergoing in vitro fertilization (IVF) treatment. More particularly, the present invention relates to a method of growing an embryo to a blastocyst stage of development comprising the step of coculturing said embryo in the presence of a population of cumulus cells. 1. A method of growing an embryo to a blastocyst stage of development comprising the step of coculturing said embryo in the presence of a population of cumulus cells.2. The method according to wherein said embryo is cultured on a cell culture surface coated with a layer of cumulus cells.3. The method according to wherein the cumulus cells are previously treated to stop their proliferation before contacting the embryo.4. The method according to wherein said cumulus cells are derived from a cumulus cell line.5. The method according to wherein said embryo and cumulus cells are human.6. The method according to wherein said embryo is generated in vitro by a technique selected from the group consisting of in vitro fertilization (IVF) claim 1 , intracytoplasmic injection of sperm into an oocyte claim 1 , and nuclear transfer.7. A method of maintaining the undifferentiated state in culture of a population of pluripotent stem cells comprising the step of coculturing said population of pluripotent stems cells in the presence of a population of cumulus cells.8. The method according to wherein said population of pluripotent stem cells is cultured on a cell culture surface coated with a layer of cumulus cells.9. The method according to wherein the cumulus cells are previously treated to stop their proliferation before contacting the pluripotent stem cells.107. The method according to wherein said cumulus cells are derived from a cumulus cell line. ...

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Номер записи: 30
13-06-2013 дата публикации

METHOD OF IN-VITRO FERTILIZATION WITH SPERMATOZOA SEPARATED INTO X-CHROMOSOME AND Y-CHROMOSOME BEARING POPULATIONS

Номер: US20130149784A1
Автор: Lu Kehuan, Seidel George E., Suh Tae Kwang
Принадлежит: XY, LLC

An IVF system for successfully utilizing spermatozoa separated into X-chromosome bearing and into Y-chromosome bearing population for insemination. The IVF system includes fertilization medium that can shorten the time from insemination to cleavage and a portable incubator for the transportation of maturing oocytes and inseminated oocytes comprising a straw () and an incubation element () that can be sealed with a cap (). 121-. (canceled)22. A method of in vitro fertilization of oocytes comprising:a) obtaining oocytes of a non-human species of mammal;b) obtaining sex sorted sperm cells of said non-human species of mammal;c) transferring a plurality of oocytes to a fertilization medium containing both essential and nonessential amino acids; andd) fertilizing at least one of the plurality of oocytes in said fertilization media with the sex sorted sperm.23. The method of claim 22 , wherein the fertilization medium having essential amino acids and non-essential amino acids improves the embryo quality of oocytes fertilized with sex sorted sperm.24. The method of claim 22 , wherein said fertilization medium is selected from the group consisting of: chemically defined medium claim 22 , SOF claim 22 , TALP supplemented with Eagles Medium claim 22 , and combinations thereof.25. The method of claim 22 , wherein said female mammal is selected from the group consisting of: primates claim 22 , humans claim 22 , bovids claim 22 , ovids claim 22 , equids claim 22 , swine claim 22 , and dolphins.26. The method of claim 22 , wherein said nonessential amino acids have a concentration in said fertilization medium and wherein the fertilization medium is further supplemented with essential amino acids.27. The method of claim 26 , wherein the further supplementation with essential amino acids comprises the addition of bovine serum albumin.28. The method of claim 22 , wherein the step of transferring a plurality of oocytes to a fertilization medium further comprises transferring about 10 ...

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Номер записи: 31
20-06-2013 дата публикации

METHOD OF LIQUID NITROGEN SURFACE VITRIFICATION

Номер: US20130157362A1
Автор: Du Fuliang, Moreno Juan, Xu Jie
Принадлежит: INGURAN, LLC

A method of liquid nitrogen surface vitrification requiring an embryo washed in a rinsing medium, then incubated in a base medium and incubated in a hold medium before being washed in a vitrification medium and produced into a vitrification droplet (). For forming the droplet, vitrification medium (), an intermediary fluid such as air, followed by vitrification medium containing at least one embryo () are aspirated into the channel. The vitrification droplet consequently can contain an air bubble (). The vitrification droplet can be produced from an instrument with a channel and dropped directly into liquid phase nitrogen producing a vitrified droplet. The vitrified droplet can be stored in cryo-vessels, and warmed for revitalization of biological function of vitrified biological cell mass or tissues, such as oocytes and/or embryos. 1. A method of forming a vitrification droplet comprising the step of:a) aspirating vitrification medium into a channel;b) aspirating an intermediary fluid into the channel;c) aspirating vitrification medium containing at least one embryo or at least one oocyte into the channel; andd) expelling the vitrification medium, the air, and the vitrification medium containing the at least one embryo or at least one oocyte from the channel to form a vitrification droplet at the end of the channel.2. The method according to claim 1 , wherein the vitrification medium and the vitrification medium containing at least one embryo are separated by the intermediary fluid in the channel prior to the step of expelling.3. The method according to claim 1 , wherein the step of expelling vitrification medium claim 1 , the intermediary fluid claim 1 , and the vitrification medium containing the at least one embryo or at least one oocyte from the channel results in a vitrification droplet formed at the end of the channel entraining the at least one embryo or at least one oocyte.4. The method according to claim 1 , wherein the intermediary fluid comprises air or ...

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Номер записи: 32
20-06-2013 дата публикации

SPERM CELL PROCESSING SYSTEMS

Номер: US20130158341A1
Автор: "OBrien Justice K.", Evans Gareth, Hollinshead Fiona K., Maxwell William M.C.
Принадлежит: XY, LLC

Semen and sperm cell processing and preservation systems, and methods of producing a mammal and methods of producing mammalian embryos are disclosed. The present invention is directed to sperm cell preservation, fertilization, and insemination, maintaining or enhancing sperm quality and addressing one or more sperm cell characteristics, such as viability, motility, functionality, fertilization rates, and pregnancy rates. Further, sperm cell characteristics may be addressed within the context of various collection, handling, separation, storage, transportation, usage, fertilization, or insemination techniques. 153-. (canceled)54. A method of producing a non-human mammalian embryo comprising:obtaining cryopreserved sperm cells;thawing the cryopreserved sperm cells;sorting the thawed sperm cells;cryopreserving the sorted sperm cells;thawing the twice cryopreserved sperm cells;fertilizing at least one egg with the thawed, twice cryopreserved sperm cells; andproducing a non-human mammalian embryo.55. The method of claim 54 , further comprising the step of producing a non-human mammal from the at least one fertilized egg.56. The method of claim 54 , further comprising the step of inseminating at least one female of a species of the non-human mammal said thawed claim 54 , twice cryopreserved sperm cells.57. The method of claim 54 , wherein the step of inseminating a female comprises inseminating a superovulated female with the thawed claim 54 , twice cryopreserved sperm cells.58. The method of claim 54 , wherein the step of fertilizing further comprises fertilizing the at least one egg in vitro.59. The method of claim 54 , further comprising the step of transferring the at least one fertilized egg to a recipient female.60. The method of claim 54 , further comprising the step of obtaining at least one egg of a superovulated female of a species of the mammal and wherein the step of fertilizing includes fertilizing the at least one egg in vitro.61. The method of claim 60 , ...

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Номер записи: 33
27-06-2013 дата публикации

INORGANIC PYROPHOSPHATE AND USES THEREOF

Номер: US20130164732A1
Автор: Sutovsky Peter, Yi Young-Joo
Принадлежит: The Curators of the University of Missouri

The present invention provides a new and improved sperm stimulating additive comprising a certain amount of inorganic pyrophosphate (PPi). Addition of PPi in the media for human/animal in vitro fertilization (IVF) improves fertilization rate; addition of PPi in the semen extender for farm animal artificial insemination (AI) may improve pregnancy rates; furthermore, mammalian oocytes matured in vitro in a medium including PPi attain improved fertilization and developmental potential, while embryos cultured in medium supplemented with PPi have improved development to blastocyst. 1. A sperm preservation media comprising inorganic pyrophosphate (PPi).2. The sperm preservation media of claim 1 , wherein the concentration of PPi is between about 1 μM and about 200 μM.3. The sperm preservation media of claim 1 , wherein the concentration of PPi is between about 1 μM and about 20 μM.4. The sperm preservation media of claim 1 , wherein the concentration of PPi is about 10 μM.5. The sperm preservation media of claim 1 , wherein said preservation media is used to preserve sperm from a porcine.6. A media for sperm processing comprising inorganic pyrophosphate (PPi).7. The media of claim 6 , wherein the concentration of PPi is between about 1 μM and about 200 μM.8. The media of claim 6 , wherein the concentration of PPi is between about 1 μM and about 20 μM.9. A media for in vitro fertilization (IVF) or artificial insemination (AI) comprising inorganic pyrophosphate (PPi).10. The media of claim 9 , wherein the concentration of PPi is between about 1 μM and about 200 μM.11. The media of claim 9 , wherein the concentration of PPi is between about 1 μM and about 20 μM.12. A semen sexing method claim 9 , comprising:(a) separating a mixed sperm suspension in a first culture medium into a population of x-bearing or y-bearing sperm with the aid of an elutant medium;(b) preserving the x-bearing or y-bearing sperm in a second culture medium,wherein, inorganic pyrophosphate (PPi) is added ...

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Номер записи: 34
11-07-2013 дата публикации

SYNTHETIC MATRICES FOR SELF-RENEWAL AND EXPANSION OF STEM CELLS

Номер: US20130177980A1
Автор: Chang Chien-Wen, Hwang Yongsung, Varghese Shyni
Принадлежит: The Regents of the University of California

Provided herein is a synthetic polymer-based hydrogel for the self-renewal and expansion of human stem cells such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). Also provided are methods of making and using the same. 1. A synthetic hydrogel comprising a heparin mimetic moiety and an acrylamide monomer.2. The synthetic hydrogel of claim 1 , wherein the heparin mimetic moiety is poly(sodium-4-styrenesulfonate) (PSS).3. The synthetic hydrogel of claim 2 , wherein the molar fraction of PSS ranges from about 0.5 to 2.4. The synthetic hydrogel of claim 2 , wherein the PSS has a matrix rigidity of about 54 kPa claim 2 , about 138 kPa claim 2 , or about 344 kPa.5. The synthetic hydrogel of claim 3 , wherein the molar ratio of the acrylamide monomer to PSS is 6:0.5 claim 3 , 6:1 claim 3 , or 6:2.6. The synthetic hydrogel of claim 5 , wherein the molar ratio of the acrylamide monomer to PSS is 6:2.7. The synthetic hydrogel of claim 1 , further comprising a bisacrylamide.8. A method for synthesizing a hydrogel comprising copolymerizing an acrylamide monomer with an ionic monomer.9. The method of claim 8 , wherein the ionic monomer is sodium-4-styrenesulfonate (SS).10. The method of claim 8 , wherein the ionic monomer is poly(sodium-4-styrenesulfonate) (PSS).11. The method of claim 10 , wherein the molar ratio of the acrylamide monomer to PSS is 6:0.5 claim 10 , 6:1 claim 10 , or 6:2.12. The method of claim 10 , wherein the PSS has a matrix rigidity of about 54 kPa claim 10 , about 138 kPa claim 10 , or about 344 kPa.13. The method of claim 8 , further comprising copolymerizing the acrylamide monomer and the ionic monomer with a bisacrylamide.14. A method for synthesizing a hydrogel comprising dissolving an acrylamide monomer and an ionic monomer in deionized water claim 8 , and polymerizing the acrylamide monomer and ionic monomer using a crosslinker and a redox initiator.15. The method of claim 14 , wherein the crosslinker is bisacrylamide.16. The ...

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Номер записи: 35
18-07-2013 дата публикации

NATIVE WHARTON'S JELLY STEM CELLS AND THEIR PURIFICATION

Номер: US20130183273A1
Автор: Taghizadeh Rouzbeh R.
Принадлежит: Auxocell Laboratories, Inc.

Noncultured Wharton's Jelly stem cells and methods of their purification, storage and use are provided. 1. A method of purifying noncultured Wharton's Jelly stem cells , the method comprising separating noncultured Wharton's Jelly stem cells from a digested tissue comprising Wharton's Jelly.2. The method of claim 1 , further comprising digesting a tissue comprising Wharton's Jelly.3. The method of claim 2 , wherein the digestion comprises exposure of the tissue to an enzyme.4. The method of claim 3 , wherein the enzyme is a collagenase.5. The method of claim 3 , wherein the enzyme is not a collagenase.6. The method of claim 2 , further comprising mechanically increasing the surface area of the tissue.7. The method of claim 6 , comprising cutting the tissue.8. The method of claim 6 , comprising mincing the tissue.9. The method of claim 2 , further comprising removing arteries and veins from a tissue comprising Wharton's Jelly prior to digesting the tissue.10. The method of claim 2 , further comprising separating digested and undigested tissue.11. The method of claim 10 , comprising sedimenting the undigested tissue.12. The method of claim 11 , wherein centrifugation is used for sedimenting the undigested tissue.13. The method of claim 10 , comprising filtering the digested tissue.14. The method of claim 13 , wherein the noncultured Wharton's Jelly stem cells are separated from the filtrate.15. The method of claim 10 , further comprising washing or diluting the digested tissue before separating it from the undigested tissue.16. A method of purifying cultured and noncultured Wharton's Jelly stem cells claim 10 , the method comprising:{'claim-ref': {'@idref': 'CLM-00010', 'claim 10'}, "purifying noncultured Wharton's Jelly stem cells according to the method of ; and"}culturing mesenchymal stem cells from the undigested tissue.17. The method of claim 16 , wherein the mesenchymal stem cells are cultured in digested Wharton's Jelly.18. Purified claim 16 , noncultured ...

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Номер записи: 36
25-07-2013 дата публикации

Protein-Free Gamete and Embryo Handling and culture Media Products

Номер: US20130189670A1
Автор: Ali bin M. Abdullah Jaffar
Принадлежит:

This invention discloses a substantially protein-free cell culture solution for assisted reproductive technologies and methods of use thereof. 1. A substantially protein-free cell culture medium for human reproductive cells , suitable for use throughout in vitro treatment of human reproductive cells to be used in in vitro fertilization and other assisted reproduction technologies , comprising mineral salts , amino acids , antioxidants , vitamins , nutrients , antibiotics , D-mannitol , and methylcellulose that has a molecular weight of 14 ,000 Daltons.2. The substantially protein-free medium according to claim 1 , wherein said methylcellulose is characterized so that a 2% solution has a viscosity of 15 centipoise at 25° C.4. The substantially protein-free medium according to claim 3 , wherein the average number of CHsubstituents attached to each sugar moiety of the compound of formula I is 1.5 to 1.9.5. The substantially protein-free medium according to claim 4 , wherein said methylcellulose is present in the solution at a concentration from 0.01 g/L to 0.5 g/L.6. The A substantially protein-free medium according to claim 4 , wherein said methylcellulose is present in the solution at a concentration from 0.01 g/L to 0.15 g/L.7. The substantially protein-free medium according to claim 4 , wherein said methylcellulose is present in the solution at a concentration of 0.1 g/L.8. The substantially protein-free medium according to claim 4 , wherein the amino acids are L-arginine claim 4 , L-cystine claim 4 , L-histidine claim 4 , L-isoleucine claim 4 , L-leucine claim 4 , L-lysine claim 4 , L-methionine claim 4 , L-phenylalanine claim 4 , L-threonine claim 4 , L-tryptophan claim 4 , L-tyrosine claim 4 , L-valine claim 4 , L-alanine claim 4 , L-taurine claim 4 , L-glutamic acid claim 4 , L-glutamine or glycine claim 4 , or any combination thereof claim 4 , wherein the amino acids are present at concentrations between 0.018 mM and 0.18 mM for L-arginine HCl; 0.0025 mM and 0 ...

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Номер записи: 37
25-07-2013 дата публикации

COMPOSITIONS AND METHODS FOR REPROGRAMMING MAMMALIAN CELLS

Номер: US20130189741A1
Автор: Chin Cynthia, Dahl Gary, Jendrisak Jerome, Meis Judith, Person Anthony
Принадлежит: CELLSCRIPT, INC.

The present invention relates to methods for changing the state of differentiation of a eukaryotic cell, the methods comprising introducing mRNA encoding one or more reprogramming factors into a cell and maintaining the cell under conditions wherein the cell is viable and the mRNA that is introduced into the cell is expressed in sufficient amount and for sufficient time to generate a cell that exhibits a changed state of differentiation compared to the cell into which the mRNA was introduced, and compositions therefor. For example, the present invention provides mRNA molecules and methods for their use to reprogram human somatic cells into pluripotent stem cells. 1. An ex vivo method for inducing a biological or biochemical effect in cells in culture that , comprising:repeatedly or continuously, at least once per day over a plurality of days, contacting the cells in culture with an RNA composition comprising in vitro-transcribed ssRNA or mRNA that encodes at least one protein, wherein the amount of dsRNA of a size greater than about 40 basepairs is less than about 0.001% of the total RNA in said RNA composition, and culturing the cells under conditions wherein said biological or biochemical effect is induced.2. The method of claim 1 , wherein said biological effect or biochemical effect is reprogramming the cells from a first differentiated state or phenotype to a second differentiated state of phenotype.3. The method of claim 1 , wherein the in vitro-transcribed ssRNA or mRNA encodes at least one protein selected from the group consisting of:i) OCT3/4, SOX1, SOX2, SOX3, SOX15, KLF1, KLF2, KLF4, KLF5, c-MYC, L-MYC, n-MYC, cMYC(T58A), LIN28, and NANOG; orii) MYOD or functional fragment or variant thereof, MYF5 or functional fragment or variant thereof, MYOGENIN or functional fragment or variant thereof, and MRF4 (MY6) or functional fragment or variant thereof; oriii) ASCL1 or functional fragment or variant thereof, MYT1L or functional fragment or variant thereof, ...

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Номер записи: 38
25-07-2013 дата публикации

MODULATION OF THE MAMMALIAN RECEPTOR mTOR TO INHIBIT STEM CELL DIFFERENTIATION INTO NEURONS

Номер: US20130189775A1
Автор: Fang Ye, Faris Ronald Allen, pai Sadashiva Karnire, Petzold Odessa Natalie
Принадлежит:

The present application relates to compositions and methods for inhibiting the differentiation of isolated embryonic stem cells, induced pluripotent stem cells, parthenogentic stem cells, or isolated embryoid bodies into neuronal progenitor cells or neuron cells comprising a mTOR inhibitor and an effective amount of cell culture growth media. The mTOR inhibitor can be Rapamycin. 1. A composition for inhibiting the differentiation of isolated embryonic stem cells , induced pluripotent stem cells , parthenogentic stem cells , or isolated embryoid bodies into neuronal progenitor cells or neuron cells comprising an amount of an effective cell culture growth media containing a mTOR inhibitor at a concentration of about 10 nM to about 100 μM.2. The composition of containing a mTOR inhibitor at a concentration of about 500 nM to about 5 μM.3. The composition of containing a mTOR inhibitor at a concentration of about 1 μM to about 3 μM.4. The composition of wherein the mTOR inhibitor is selected from the group of rapamycin claim 1 , KU-0063794 claim 1 , Pp242 and WYE-354 claim 1 , temsirolimus claim 1 , or everolimus.5. The composition of wherein the stem cells are of mammalian origin.6. The composition of wherein the stem cells are one or more of human cells claim 5 , simian cells claim 5 , or murine cells.7. The composition of wherein the isolated embryonic stem cells are isolated human embryonic stem cells.8. An in vitro method for inhibiting the differentiation of one or more of a stem cell selected from the group of an isolated embryonic stem cell claim 1 , an induced Pluripotent stem cell claim 1 , a parthenogenetic stem cell or an isolated embryoid body into a population of neural progenitor cells and/or a neurons comprising contacting an effective amount of a composition of with the stem cell claim 1 , for an effective amount of time claim 1 , thereby inhibiting the differentiation of the stem cells into a neural progenitor cells or neurons.9. The method of claim 8 ...

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Номер записи: 39
25-07-2013 дата публикации

METHODS FOR THE PRODUCTION OF IPS CELLS USING NON-VIRAL APPROACH

Номер: US20130189778A1
Автор: MACK Amanda
Принадлежит: CELLULAR DYNAMICS INTERNATIONAL, INC.

Methods and composition of induction of pluripotent stem cells and other desired cell types are disclosed. For example, in certain aspects methods for generating essentially vector-free induced pluripotent stem cells are described. Furthermore, the invention provides induced pluripotent stem cells and desired cell types essentially free of exogenous vector elements with the episomal expression vectors to express differentiation programming factors. 1. A human iPS cell population that is essentially free of exogenous retroviral elements , the cell population comprising the genome of a selected human individual.2. A human iPS cell population comprising cells whose genome is derived from a terminally differentiated human cell and essentially free of exogenous retroviral elements.3. The iPS cell population of claim 1 , wherein the iPS cell population is essentially free of exogenous viral vector elements.439.-. (canceled)40. The iPS cell population of claim 2 , wherein the iPS cell population is essentially free of exogenous viral vector elements. This application is a divisional of U.S. application Ser. No. 12/478,154, filed on Jun. 4, 2009, which claims priority to U.S. Application No. 61/058,858, filed on Jun. 4, 2008 and U.S. Application No. 61/160,584, filed on Mar. 16, 2009. The entire text of each of the above referenced disclosures is specifically incorporated herein by reference.1. Field of the InventionThe present invention relates generally to the field of molecular biology, stem cells and differentiated cells. More particularly, it concerns differentiation programming or reprogramming of somatic cells and undifferentiated cells.2. Description of Related ArtIn general, stem cells are undifferentiated cells which can give rise to a succession of mature functional cells. For example, a hematopoietic stem cell may give rise to any of the different types of terminally differentiated blood cells. Embryonic stem (ES) cells are derived from the embryo and are ...

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Номер записи: 40
25-07-2013 дата публикации

REPROGRAMMING COMPOSITIONS

Номер: US20130189780A1
Автор: Flynn Peter, Shoemaker Daniel
Принадлежит: FATE THERAPEUTICS, INC.

The present invention provides compositions and methods of using the compositions to alter the developmental potency of a cell. The present invention provides in vivo and ex vivo cell reprogramming or dedifferentiation methods suitable for autologous cell therapy and regenerative medicine. 2. The method of claim 1 , wherein the APTF comprises a cell permeable peptide (CPP).34.-. (canceled)5. The method of claim 1 , wherein the DBD is selected from the group consisting of: Oct-3/4 claim 1 , Cdx-2 claim 1 , Gbx2 claim 1 , Gsh1 claim 1 , HesX1 claim 1 , HoxA10 claim 1 , HoxA11 claim 1 , HoxB1 claim 1 , Irx2 claim 1 , Isl1 claim 1 , Meis1 claim 1 , Meox2 claim 1 , Nanog claim 1 , Nkx2.2 claim 1 , Onecut claim 1 , Otx1 claim 1 , Oxt2 claim 1 , Pax5 claim 1 , Pax6 claim 1 , Pdx1 claim 1 , Tcf1 claim 1 , Tcf2 claim 1 , Zfhx1b claim 1 , Klf-4 claim 1 , Atbf1 claim 1 , Esrrb claim 1 , Gcnf claim 1 , Jarid2 claim 1 , Jmjd1a claim 1 , Jmjd2c claim 1 , Klf-3 claim 1 , Klf-5 claim 1 , MeI-18 claim 1 , Myst3 claim 1 , Nac1 claim 1 , REST claim 1 , Rex-1 claim 1 , Rybp claim 1 , Sall4 claim 1 , Sall1 claim 1 , Tif1 claim 1 , YY1 claim 1 , Zeb2 claim 1 , Zfp281 claim 1 , Zfp57 claim 1 , Zic3 claim 1 , Coup-Tf1 claim 1 , Coup-Tf2 claim 1 , Bmi1 claim 1 , Rnf2 claim 1 , Mta1 claim 1 , Pias1 claim 1 , Pias2 claim 1 , Pias3 claim 1 , Piasy claim 1 , Sox2 claim 1 , Lef1 claim 1 , Sox15 claim 1 , Sox6 claim 1 , Tcf-7 claim 1 , Tcf711 claim 1 , c-Myc claim 1 , L-Myc claim 1 , N-Myc claim 1 , Hand1 claim 1 , Mad1 claim 1 , Mad3 claim 1 , Mad4 claim 1 , Mxi1 claim 1 , Myf5 claim 1 , Neurog2 claim 1 , Ngn3 claim 1 , Olig2 claim 1 , Tcf3 claim 1 , Tcf4 claim 1 , Foxc1 claim 1 , Foxd3 claim 1 , BAF155 claim 1 , C/EBPβ claim 1 , mafa claim 1 , Eomes claim 1 , Tbx-3; Rfx4 claim 1 , Stat3 claim 1 , Stella claim 1 , and UTF-1.68.-. (canceled)9. The method of claim 1 , wherein the TAD is selected from the group consisting of: VP16 claim 1 , VP64 claim 1 , SV40 Large T-antigen claim 1 , E1A ...

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Номер записи: 41
01-08-2013 дата публикации

INDUCED PLURIPOTENT STEM CELLS

Номер: US20130195812A1
Автор: HIRAI HIROYUKI, Kikyo Nobuaki
Принадлежит: Regents of the University of Minnesota

Described herein is a major breakthrough in nuclear reprogramming and induced pluripotent stem cell (iPSC) technology. Fusion of the powerful transcription activation domain (TAD) of MyoD to the Oct4 protein makes iPSCs generation faster, more efficient, purer, safer and feeder-free. Also, disclosed herein is the first report of the use of a TAD fused to a transcription factor as a method for making iPSCs. By combining transcription factors and TADs, this approach to nuclear reprogramming can have a range of applications from inducing pluriopotency to inducing transdifferentiation without transitioning through iPSCs. 1. A method for preparing an induced pluripotent stem cell comprising introducing a nucleic acid sequence which is a fusion of a heterologous transactivation domain and a transcription factor into a somatic cell.2. The method of claim 1 , wherein the transcription factor is Oct-4.3. The method of claim 1 , wherein the transactivation domain is obtained from MyoD or VP16.4. The method of claim 3 , wherein the transactivation domain of MyoD comprises an N-terminus region of MyoD.5. The method of claim 4 , wherein the transactivation domain comprising amino acids 1-62 of MyoD or is at least 80% identical thereto.6. The method of claim 1 , wherein a transcription factor is further introduced into the cell.7. The method of any one of claim 6 , wherein the transcription factor is Klf4.8. The method of any one of claim 6 , wherein transcription factor is Sox2 claim 6 , c-Myc or a combination thereof.9. The method of claim 1 , wherein the cell is mammalian.10. The method of claim 9 , wherein the mammalian cell is human.11. A fusion nucleic acid or protein comprising a heterologous transactivation domain and a transcription factor.12. The fusion of claim 11 , wherein the transcription factor is Oct4.12. The fusion of claim 11 , wherein the transactivation domain (TAD) is obtained from MyoD or VP16.13. The fusion of claim 11 , wherein the transactivation domain ( ...

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Номер записи: 42
01-08-2013 дата публикации

TISSUE-SPECIFIC DIFFERENTIATION MATRICES AND USES THEREOF

Номер: US20130195814A1
Автор: Chen Xiao-Dong
Принадлежит: The Board of Regents of the University of Texas System

In some aspects, this invention provides a method of making a bone marrow-derived tissue-specific stem cell proliferation, expansion, isolation and rejuvenation extracellular matrix. In other aspects, this invention provides a method of making a tissue-specific fibroblast-derived stem cell differentiation extracellular matrix. Also provided are methods of using such a cell-derived preservation or differentiation matrices to induce tissue-specific differentiation of pluripotent cells, repair damaged tissue, and treat a subject having a physiologic deficiency using the same. 153-. (canceled)54. A tissue-specific differentiation matrix comprising an extracellular matrix generated by target tissue-specific fibroblast cells.55. The tissue-specific differentiation matrix of claim 54 , wherein the fibroblast cells are human or mouse fibroblast cells.56. The tissue-specific differentiation matrix of claim 54 , wherein the target tissue-specific fibroblast cells are from neural tissue claim 54 , epidermal tissue claim 54 , dermal tissue claim 54 , adipose tissue claim 54 , cardiac tissue claim 54 , kidney tissue claim 54 , muscle tissue claim 54 , liver tissue claim 54 , cartilage tissue claim 54 , pancreas tissue claim 54 , tissue of the endometrium of uterus claim 54 , umbilical cord tissue claim 54 , dental pulp tissue claim 54 , trabecular or cortical bone tissue.57. The tissue-specific differentiation matrix of claim 54 , wherein the tissue-specific differentiation matrix is a 3D tissue-specific differentiation matrix.58. The tissue-specific differentiation matrix of claim 54 , wherein the tissue-specific differentiation matrix is essentially free of feeder cells.59. The tissue-specific differentiation matrix of claim 54 , wherein the tissue-specific differentiation matrix is essentially free of fibroblast cells.60. A method of making a tissue-specific differentiation matrix comprising:a) culturing target tissue-specific fibroblast cells on a surface to produce an ...

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Номер записи: 43
01-08-2013 дата публикации

METHOD FOR SELECTING AN IPS CELL

Номер: US20130196865A1
Автор: Hochedlinger Konrad, Stadtfeld Matthias
Принадлежит: The General Hospital Corporation

This application relates to a method for selecting an induced pluripotent stem cell (iPS), the method comprising: selecting an iPS cell that expresses a gene in the Dlk1-Dio3 cluster from a population of iPS cells. The method further comprises: comparing the gene expression profile determined for an iPS cell with the gene expression profile determined for an embryonic stem cell; identifying a gene that is differentially expressed in the embryonic stem cell as compared to the iPS cell; and selecting the desired iPS cell from a population of iPS cells. 1. A method for selecting an induced pluripotent stem cell (iPS) , the method comprising: selecting an iPS cell that expresses a gene in the Dlk1-Dio3 cluster from a population of iPS cells.2. The method of claim 1 , wherein the gene is Meg3 claim 1 , Rian or Mirg.3. The method of claim 1 , wherein expression of each of genes Meg3 claim 1 , Rian and Mirg are measured.4. The method of claim 1 , wherein the induced pluripotent stem cell is a mammalian iPS cell.5. The method of claim 1 , further comprising differentiating the iPS cell selected in .6. The method of claim 1 , wherein the iPS cell expressing the identified gene in the Dlk1-Dio3 cluster has an enhanced differentiation potential compared to an iPS cell lacking expression of the identified gene in the Dlk1-Dio3 cluster.719-. (canceled)20. A method for screening for an agent that enhances iPS cell differentiation potential claim 1 , the method comprising:(a) providing an iPS cell population lacking expression of one or more genes in the Dlk1-Dio3 cluster,(b) contacting the iPS cell population with a candidate agent;(c) measuring the level of expression of the one or more genes in the Dlk1-Dio3 cluster, wherein expression of the one or more genes is indicative that the agent enhances iPS cell differentiation potential.21. The method of claim 20 , wherein the one or more genes is Meg3 claim 20 , Rian or Mirg.22. (canceled)23. The method of claim 20 , wherein the ...

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Номер записи: 44
01-08-2013 дата публикации

COMPOSITIONS AND METHODS FOR INDUCING CELL DEDIFFERENTIATION

Номер: US20130196989A1
Автор: Chen Shuibing, Ding Sheng, Schultz Peter G.
Принадлежит: The Scripps Research Institute

The present invention provides compounds, compositions and methods for dedifferentiating lineage committed mammalian cells into stem cells. The present invention also provides methods of inducing dedifferentiation of lineage committed mammalian cells into stem cells, which can be further differentiated into various lineage committed cells. Methods of identifying additional compounds useful for inducing dedifferentiation of lineage committed cells into stem cells are also provided. 19.-. (canceled)11. The compound of claim 10 , wherein Ris hydrogen claim 10 , and Ris Ccycloalkyl.12. The compound of claim 11 , wherein Ris cyclohexyl.16. A pharmaceutical composition comprising a compound of and a pharmaceutically acceptable carrier.17. The pharmaceutical composition of claim 16 , wherein Ris hydrogen claim 16 , and Ris Ccycloalkyl.18. The pharmaceutical composition of claim 17 , wherein Ris cyclohexyl.22. A method of inducing dedifferentiation of a lineage committed cell ex vivo claim 17 , the method comprising:{'claim-ref': {'@idref': 'CLM-00010', 'claim 10'}, 'contacting a lineage committed mammalian cell with a compound of , whereby the mammalian cell dedifferentiates into a multipotent stem cell.'}23. The method of claim 22 , further comprising detecting dedifferentiation of the mammalian cell into a multipotent stem cell.24. The method of claim 22 , whereby differentiation of the lineage committed mammalian cell into a multipotent stem cell is detected by detecting loss of expression of a marker gene expressed by the lineage committed mammalian cell.25. The method of claim 24 , wherein said lineage committed cell is a myoblast cell.26. The method of claim 25 , wherein the marker gene is a member selected from the group consisting of: MyoD claim 25 , Myf5 claim 25 , myosin claim 25 , CD56 and desmin.27. The method of claim 25 , wherein the myoblast cell is isolated from a mouse.28. The method of claim 25 , wherein the myoblast cell is isolated from a primate.29. ...

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Номер записи: 45
08-08-2013 дата публикации

Swellable (meth)acrylate surfaces for culturing cells in chemically defined media

Номер: US20130203165A1
Автор: Andrei Gennadyevich Fadeev, Arthur Winston Martin, Christopher Bankole Shogbon, David Michael Weber, Jennifer Lynn WEBER, Yue Zhou, Zara Melkoumian
Принадлежит: Individual

Synthetic surfaces capable of supporting culture of undifferentiated human embryonic stem cells in a chemically defined medium include a swellable (meth)acrylate layer and a peptide conjugated to the swellable (meth)acrylate layer. The swellable (meth)acrylate layer may be formed by polymerizing monomers in a composition that includes hydroxyethyl methacrylate, 2-carboxyehylacrylate, and tetra(ethylene glycol)dimethacrylate. The conjugated peptide may include an amino acid sequence of Xaa n ProGlnValThrArgGlyAspValPheThrMetPro, where n is an integer from 0 to 3 and where Xaa is any amino acid. Further, disclosed herein is a swellable (meth)acrylate synthetic surface which can be sterilized by gamma irradiation.

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Номер записи: 46
08-08-2013 дата публикации

CONDITIONED CELL CULTURE MEDIUM COMPOSITIONS AND METHODS OF USE

Номер: US20130203169A1
Автор: Applegate Mark, Horwitz David L., Kern Andreas, Landeen Lee K., Mansbridge Jonathan, Naughton Gail K., Pinney R. Emmett, Ratcliffe Anthony, Zeltinger Joan
Принадлежит: ALLERGAN, INC.

Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder. Additionally, the medium is formulated with a pharmaceutically acceptable carrier as a vehicle for internal administration, applied directly to a food item or product, or formulated with a salve or ointment for topical applications. Also, the medium may be further processed to concentrate or reduce one or more factors or components contained within the medium. 1. A medium for making a composition suitable for topical application , the medium made by a process comprising the steps of:(a) inoculating fetal or embryonic fibroblasts onto a substrate in a nutrient medium;(b) culturing the fetal or embryonic fibroblasts in a two dimensional culture until the fetal or embryonic fibroblasts secrete a desired level of a growth factor and a collagen; and(c) removing the medium from the cultured cells.2. The medium for making a composition suitable for topical application of claim 1 , wherein the growth factor is keratinocyte growth factor.3. The medium for making a composition suitable for topical application of claim 1 , wherein the substrate comprises beads.4. The medium for making a composition suitable for topical application of claim 1 , wherein during the culturing step the fetal or embryonic fibroblasts are subjected to hypoxic stress to thereby increase the secretion of the growth factor.5. The medium for making a composition suitable for topical application of claim 2 , wherein during the culturing step the fetal or embryonic fibroblasts are subjected to hypoxic stress to ...

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Номер записи: 47
15-08-2013 дата публикации

CONDITIONED CELL CULTURE MEDIUM COMPOSITIONS AND METHODS OF USE

Номер: US20130210725A1
Автор: Applegate Mark, Horwitz David L., Kern Andreas, Landeen Lee K., Mansbridge Jonathan, Naughton Gail K., Pinney R. Emmett, Ratcliffe Anthony, Zeltinger Joan
Принадлежит: ALLERGAN, INC.

Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder. Additionally, the medium is formulated with a pharmaceutically acceptable carrier as a vehicle for internal administration, applied directly to a food item or product, or formulated with a salve or ointment for topical applications. Also, the medium may be further processed to concentrate or reduce one or more factors or components contained within the medium. 1. A composition comprising a cell culture media formed by:(a) inoculating fetal or embryonic fibroblasts onto a substrate in a nutrient medium;(b) culturing the fetal or embryonic fibroblasts in a two dimensional culture until the fetal or embryonic fibroblasts secrete a desired level of a growth factor and a collagen; and(c) removing the medium from the cultured cells; wherein the composition comprises the removed medium.2. The composition of claim 1 , wherein the growth factor is keratinocyte growth factor.3. The composition of claim 1 , wherein the substrate comprises beads.4. The composition of claim 1 , wherein during the culturing step the fetal or embryonic fibroblasts are subjected to hypoxic stress to thereby increase the secretion of the growth factor.5. The composition of claim 2 , wherein during the culturing step the fetal or embryonic fibroblasts are subjected to hypoxic stress to thereby increase the secretion of the keratinocyte growth factor.6. The composition of claim 1 , further comprising the step of concentrating the medium removed from the cultured cells.7. The composition of claim 1 , further ...

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Номер записи: 48
15-08-2013 дата публикации

FEEDER-FREE METHOD FOR CULTURE OF BOVINE AND PORCINE SPERMATOGONIAL STEM CELLS

Номер: US20130211186A1
Автор: Oatley Jon Michael, Oatley Melissa Joan
Принадлежит: Washington State University Research Foundation

The present invention relates to the production and culture of undifferentiated spermatogonial stem cells that can be maintained long term and are feeder free. The resultant feeder-free populations can be used in any of a number of protocols including the generation of progeny bulls. The present invention includes novel methods required for the successful enrichment of bovine spermatogonial stem cells, novel cell lines and other components used for the same, as well as the resultant stem cell compositions. 1. A method of enriching spermatogonial stem cells (SSCs) from a population of testis-derived cells containing at least one SSC , said method comprising:a) providing a media that has been preconditioned with a feeder cell line;b) contacting said population of testis-derived cells with said preconditioned media under conditions suitable for SSC cell maintenance and enrichment.2. The method of wherein said preconditioned media is created by contacting said media with cells claim 1 , or parts thereof from a strain of bovine embryonic fibroblast cells and/or bovine somatic cells isolated from bovine testes.3. The method of wherein said cell strain is selected from the group consisting of cell line BEF1 and cell line BSC1.4. The method of wherein preconditioning includes the steps of: a) contacting media with feeder cells selected from the group consisting of bovine embryonic fibroblast cells and/or bovine testicular somatic cells; b) allowing said cells to reproduce on said media; and thereafter c) removing said feeder cells.5. The method of wherein said cell feeder cells are derived from cell line BEF1 and/or BSC1.6. The method of wherein said cells are bovine cells.7. A bovine embryonic fibroblast cell line for pre-conditioning media for culture of bovine spermatogonial stem cells BEF1 deposited as ATCC Accession number ______.8. A bovine somatic cell line for pre-conditioning media for culture of bovine spermatogonial stem cells BSC1 deposited as ATCC Accession ...

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Номер записи: 49
22-08-2013 дата публикации

PLACENTA-DERIVED CELL-CONDITIONED CULTURE MEDIA AND ANIMAL-FREE, FEEDER-FREE METHOD FOR CULTURING STEM CELLS USING THE SAME

Номер: US20130217120A1
Автор: JUNG Ji Hye Hye, KIM Byung Soo, KIM Ji Hye Hye, LEE Seung-Jin, PARK Yong
Принадлежит: KOREA UNIVERSITY RESEARCH AND BUSINESS FOUNDATION

Disclosed are placenta-derived cell-conditioned culture media for stem cells. An animal-free, feeder-free method using the media is also provided for culturing stem cells. The media can prevent the stem cells from being contaminated with xenogeneic proteins or cells, and maintain human embryonic stem cells in an undifferentiated state for a long period of time in vitro with an economic benefit. 1. A method for preparing a placenta-derived cell-conditioned culture medium , comprising:(a) seeding placenta-derived cells onto a solid surface;(b) bring a cell culture medium to be in contact with the placenta-derived cells and incubating the placenta-derived cells; and(c) recovering only the cell culture medium from the culture obtained in (b) to obtain the placenta-derived cell-conditioned culture medium,wherein said placenta-derived cell-conditioned culture medium is suitable for culturing stem cells.2. The method of claim 1 , wherein the placenta-derived cells of step (a) are placenta-derived fibroblast-like cells separated from a human chorionic plate.3. The method of claim 1 , wherein the basal cell culture medium of step (b) is DMEM/F-12 supplemented with a serum replacement.4. The method of claim 1 , wherein the incubation of step (b) is conducted for 20-30 hours.5. The method of claim 1 , wherein the solid surface in step (a) is a gelatin-coated surface.6. The method of claim 1 , wherein the placenta-derived cell-conditioned medium comprises b-FGF claim 1 , IL-8 claim 1 , TIMP-2 claim 1 , MCP-1 claim 1 , GRO and GRO-α claim 1 , and wherein the amount of b-FGF is smaller than the amount of each of IL-8 claim 1 , TIMP-2 claim 1 , MCP-1 claim 1 , GRO and GRO-α.7. The method of claim 6 , wherein the placenta-derived cell conditioned medium further comprises at least one cytokine selected from the group consisting of osteoprotegerin claim 6 , uPAR claim 6 , TIMP-1 claim 6 , IGFBP-6 claim 6 , ICAM-1 claim 6 , angiogenin claim 6 , BDNF claim 6 , IGFBP-4 claim 6 , IGFBP-2 ...

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Номер записи: 50
29-08-2013 дата публикации

LARGE-SCALE PROPAGATION AND MAINTENANCE METHOD OF EMBRYOID BODIES GENERATED FROM STEM CELLS

Номер: US20130224856A1
Автор: Cho Yee Sook, Kim Hyun Jin, Son Mi Young
Принадлежит: Korea Research Institute of Bioscience and Biotechnology

The present invention relates to a large-scale propagation and maintenance method of embryoid bodies generated from stem cells. More particularly, the present invention relates to a large-scale propagation and maintenance method of embryoid bodies retaining their intrinsic characteristics for a long period of time, comprising the step of continuously subculturing embryoid bodies that are primarily produced from embryonic stem cells or from induced pluripotent stem cells. According to the method of the present invention, after preparation of embryoid bodies from a limited number of stem cells, large-scale production and maintenance of embryoid bodies can be realized by a simple mechanical subculturing method without the continuous supply of stem cells. 1. A large-scale propagation method of embryoid bodies , comprising the step of subculturing embryoid bodies generated from stem cells.2. The method according to claim 1 , wherein the stem cells are pluripotent stem cells.3. The method according to claim 1 , wherein the stem cells are embryonic stem cells or induced pluripotent stem cells.4. The method according to claim 1 , wherein the stem cell are derived from human.5. The method according to claim 1 , wherein the subculturing is performed by mechanical or enzymatic division.6. The method according to claim 5 , wherein the mechanical division is performed using a blade claim 5 , a tissue chopper claim 5 , a needle claim 5 , a pipette claim 5 , an EBD (embryoid body divider) or a scraper.7. The method according to claim 5 , wherein the enzymatic division is performed using collagenase claim 5 , accutase claim 5 , dispase or trypsin.8. The method according to claim 1 , wherein the step of subculturing embryoid bodies is performed by suspension culture.9. The method according to claim 1 , wherein the embryoid bodies are established by suspension culture of colonies of collected stem cells cut into the dimension of 10 to 1000 μm on all sides claim 1 , undisturbed ...

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Номер записи: 51
05-09-2013 дата публикации

Renovation And Repopulation Of Decellularized Tissues And Cadaveric Organs By Stem Cells

Номер: US20130230497A1
Автор: Hariri Robert J.
Принадлежит: Anthrogenesis Corporation

A method of manufacturing a tissue matrix for implantation into a patient is disclosed. The method sets forth collecting embryonic stem cells from a placenta which has been treated to remove residual cord blood and seeding the collected stem cells onto or into a tissue matrix. The seeded tissue matrix is then implanted on or into a patient. The seeded tissue matrix made by the method of the present invention is also disclosed. 1. A method of manufacturing a tissue matrix for implantation into a patient , comprising: collecting embryonic stem cells from a placenta which has been treated to remove residual cord blood and seeding said collected stem cells onto or into a tissue matrix for implantation into a patient.2. The method of further comprising perfusing a placenta which has been drained of cord blood with an anticoagulant solution to flush out residual cells and collecting said residual cells and perfusion liquid from the drained placenta for seeding into or onto the tissue matrix.3. The method of further comprising separating said embryonic stem cells from said residual cells and perfusion liquid prior to seeding the tissue matrix.4. The method of including wherein the placenta is perfused with the anticoagulant solution by passing the anticoagulant solution into one or both of the umbilical artery and umbilical vein.5. The method of including wherein said residual cells and perfusion liquid are collected from the part of the placenta that was attached to the wall of the uterus of the mother.6. The method of including wherein said embryonic stem cells are separated from said residual perfusion liquid by centrifugation.7. The method of including wherein the tissue matrix is a decellularized tissue.8. The method of including wherein said tissue matrix is washed to assure removal of cellular and extracellular debris prior to said seeding.9. The method of including wherein said tissue matrix prior to said seeding is treated with factors to enhance the adhesion of ...

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Номер записи: 52
05-09-2013 дата публикации

Equine Amniotic Fluid-Derived Multipotent Stem Cells and a Method for Producing the Same

Номер: US20130230924A1
Автор: Kang Kyung Sun, PARK Sang Bum, SEO Min Soo
Принадлежит: KANG STEM HOLDINGS CO., LTD

The present invention relates to equine amniotic fluid-derived multipotent stem cells (eAF-MSCs) and a preparation method thereof. More particularly, the present invention relates to equine amniotic fluid-derived multipotent stem cells which exhibit all negative immunological characteristics with respect to the human markers, CD19, CD20, CD28, CD31, CD34, CD38, CD41a, CD62L, CD62P and CD200, and exhibit all positive immunological characteristics with respect to the human markers, CD44, CD90 and CD105, and have the ability to differentiate into ectoderm, mesoderm or endoderm-derived cells. 1. A method for preparing equine amniotic fluid-derived multipotent stem cells showing the following characteristics and being more homogeneous than those before culture , comprisingStep 1 of culturing stem cells isolated from equine amniotic fluid in EGM-2 (Endothelial cell Growth Medium-2); andStep 2 of recovering the cultured cells:(a) showing all negative immunological characteristics with respect to the human markers CD19, CD20, CD28, CD31, CD34, CD38, CD41a, CD62L, CD62P and CD200, and showing all positive immunological characteristics with respect to the human markers CD44, CD90 and CD105;(b) having the ability to differentiate into ectoderm, mesoderm or endoderm-derived cells; and(c) having the ability to maintain an undifferentiated state for 14 passages or more.2. The method according to claim 1 , wherein the stem cells isolated from equine amniotic fluid in Step 1 are those isolated from a middle layer claim 1 , cell layer claim 1 , after centrifugation using a density-gradient solution.3. The method according to claim 1 , wherein the culturing of Step 1 is culturing of the stem cells isolated from equine amniotic fluid in attached plate with EGM-2 (Endothelial cell Growth Medium-2).4. The method according to claim 1 , wherein fetal bovine serum (FBS) is added to EGM-2 in Step 1.5. The method according to claim 1 , wherein the cells cultured in Step 1 are further ...

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Номер записи: 53
12-09-2013 дата публикации

Reprogramming of Aged Adult Stem Cells

Номер: US20130236428A1
Автор: Giampapa Vincent C.
Принадлежит:

Reprogramming of mammalian stem cells including aged human adult stem cells of all types with young adult stem cell's supernatant-intracellular matrix, bioactive lipids and or microvesicules in a single step or as a secondary two step process using oocyte supernatant, its intracellular matrix and/or cellular components are disclosed to accomplish a gene erasure and reprogramming. This invention focuses on reprogramming and/or reactivating genes that are active and involved in youthful adult stem cell function, within aged adult stem cells that have been previously collected by and/or stored for patients who are 40 years and older. The process is accomplished by using the natural unaltered young adult stem cell fluid and its cellular components. 1. A method of reprogramming aged human or animal adult stem cells (AASC) of all types , comprising the steps of:(a) collecting young adult stem cells (YASC) from the blood of a donor, using aphesis or like means;(b) using cell lysis or simple diffusion to at least partially dissolve membranes of said YASC to release an intracellular matrix (ICM) thereof;(c) applying a supernatant of said ICM of said YASC to a culture of AASC to be reprogrammed; and(d) following exposure of said AASC to said supernatant for a bioactively sufficient period, infusing such an exposed AASC to the donor thereof.2. The method as recited in claim 1 , further comprising:re-iterating said Steps (b) and (c).3. The method as recited in claim 1 , further comprising:separating said YASC from said supernatant of said YASC by use of a membrane and permitting YASC soluble factors and related components to pass therethrough prior to application of said Step (c).4. The method as recited in claim 3 , further comprising:re-iterating said Steps (b) and (c).5. The method as recited in claim 1 , further comprising the steps of:(e) prior to said Step (a), collecting oocytes biologically compatible with said AASC to be re-programmed;(f) using lysis or like means to ...

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Номер записи: 54
12-09-2013 дата публикации

VITRIFICATED STORAGE SOLUTION FOR CELLS

Номер: US20130236960A1
Автор: Kunitomi Yoshihiro, Sasaki Masahiro
Принадлежит:

The vitrification medium for cells according to the present invention is a vitrification medium for cells comprising a cell membrane permeable substance and a cell membrane non-permeable substance, in which, the content of the cell membrane permeable substance is in the range of 30 to 50% by volume, and the osmotic pressure generated in association with the cell membrane non-permeable substance, which is a fraction of the total osmotic pressure on the cell membrane on suspending the cells in the vitrification medium, is in the range of 280 mOsm or more. The vitrification medium for cells according to the present invention is excellent in operability and safety. 1. A vitrification medium for cells comprising a cell membrane permeable substance and a cell membrane non-permeable substance , in whichthe content of the cell membrane permeable substance is in the range of 30 to 50% by volume, andthe osmotic pressure generated in association with the cell membrane non-permeable substance, which is a fraction of the total osmotic pressure on the cell membrane on suspending the cells in the vitrification medium, is in the range of 280 mOsm or more.2. A vitrification medium for cells according to claim 1 , wherein the cell membrane permeable substance is one or more selected from the group consisting of ethylene glycol (EG) claim 1 , propylene glycol (PG) claim 1 , 1 claim 1 ,3-propanediol (1 claim 1 ,3-PD) claim 1 , butylene glycol (BG) claim 1 , isoprene glycol (IPG) claim 1 , dipropylene glycol (DPG) claim 1 , glycerin and dimethyl sulfoxide (DMSO).3. A vitrification medium for cells according to claim 1 , wherein the cell membrane permeable substance is one or two selected from the group consisting of ethylene glycol (EG) claim 1 , propylene glycol (PG) and dimethyl sulfoxide (DMSO).4. A vitrification medium for cells according to claim 1 , wherein the cell membrane non-permeable substance is one or more selected from the group consisting of sodium chloride claim 1 , ...

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Номер записи: 55
12-09-2013 дата публикации

Medium and Culture of Embryonic Stem Cells

Номер: US20130236962A1
Автор: Ludwig Tenneille, Thomson James A.
Принадлежит: WISCONSIN ALUMNI RESEARCH FOUNDATION

Previous methods for culturing primate pluripotent stem cells have required either fibroblast feeder cells or a medium which was exposed to fibroblast feeder cells to maintain the stem cells in an undifferentiated state. It has now been found that high levels of fibroblast growth factor in a medium together with at least one of gamma aminobutyric acid, pipecolic acid, and lithium, enables pluripotent stem cells to remain undifferentiated indefinitely through multiple passages, even without feeder cells or conditioned medium. Without beta-mercaptoethanol, the medium improves cloning efficiency. Also, a matrix of human proteins can be used to culture the undifferentiated cells without exposing the cells to animal products. Further disclosed are new primate pluripotent cell lines made using the defined culture conditions, including the medium and the matrix. Such new cell lines will have never been exposed to animal cells, animal products, feeder cells or conditioned medium. 1. A method for culturing primate pluripotent stem cells in an undifferentiated state on a matrix without the need for feeder cells or conditioned medium , the method comprising the step of:culturing the primate pluripotent stem cells on a matrix in a medium without feeder cells or conditioned media, the medium comprising salts, vitamins, amino acids, glucose, albumin, minerals, lipids, a transferrin or a transferrin substitute, insulin or an insulin substitute, a fibroblast growth factor, and at least one member selected from gamma-aminobutyric acid, pipecolic acid, and lithium in sufficient amounts to maintain the cells in an undifferentiated state through multiple successive culture passages.2. The method of wherein the matrix comprises a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm mouse sarcoma or a matrix that comprises human matrix proteins collagen IV and at least one member selected from fibronectin claim 1 , laminin claim 1 , and vitronectin.3. The ...

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Номер записи: 56
12-09-2013 дата публикации

Defined Media for Expansion and Maintenance of Pluripotent Stem Cells

Номер: US20130236973A1
Автор: Rezania Alireza
Принадлежит: JANSSEN BIOTECH, INC.

The present invention provides methods to promote the proliferation of undifferentiated pluripotent stem cells in defined media. Specifically, the invention provides a defined cell culture formulation for the culture, maintenance, and expansion of pluripotent stem cells, wherein culturing stem cells in the defined cell culture formulation maintains the pluripotency and karyotypic stability of the cells for at least 10 passages. Further disclosed is a cell population grown under defined media conditions that express OCT4, SOX2, NANOG, and FOXA2. 1. A defined cell culture formulation for the culture , maintenance , and expansion of pluripotent stem cells , wherein the defined cell culture formulation comprises basal medium , insulin , transferrin , selenium , fatty-acid free albumin , a TGF-β ligand , bFGF , and ascorbic acid; and wherein culturing stem cells in the defined cell culture formulation maintains the pluripotency and karyotypic stability of the cells for at least 10 passages.2. The defined cell culture formulation of claim 1 , wherein the cell culture formulation further comprises insulin growth factor 1 (IGF-1).3. The defined cell culture formulation of or claim 1 , wherein the cell culture formulation comprises DMEM-F12.4. The defined cell culture formulation of claim 1 , wherein the cell culture formulation further comprises Trace Elements C claim 1 , 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid claim 1 , lithium chloride claim 1 , glucose claim 1 , Defined Lipids claim 1 , and L-alanyl-L-glutamine dipeptide.5. The defined cell culture formulation of claim 4 , wherein the cell culture formulation comprises MCDB-β1.6. The defined cell culture formulation of any one of to claim 4 , wherein ITS-X provides the insulin claim 4 , transferrin claim 4 , and selenium.7. The defined cell culture formulation of any one of to claim 4 , wherein the fatty acid free albumin is reagent grade.8. The defined cell culture formulation of any one of to claim 4 , ...

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Номер записи: 57
26-09-2013 дата публикации

Device for performing micro-operations on a vesicular object

Номер: US20130252318A1
Автор: Taylor Paul J.
Принадлежит:

A device for performing micro-operations on a vesicular target, comprising an injection pipette (); a barrel assembly (); and an outer assembly (). The injection pipette, barrel assembly, and outer assembly being designed and situated in relation to each other such that an axial vacuum passage is created between the injection pipette and the barrel assembly, and a radial vacuum passage is created between the barrel assembly and outer assembly, and such vacuum passages are isolated from each other and from atmospheric pressure (FIG. ). The device also comprises a means of advancing and withdrawing the distal end of the barrel assembly in relation to the distal end of the outer assembly so as to create a holding well () and a means of advancing the pipette into, and withdrawing the pipette from, a vesicular object. The elements being designed and situated in relationship to each other to allow the simultaneous holding of the vesicular object by applying negative pressure in the radial vacuum passage; injection into or aspiration from the vesicular object via the pipette; and/or aspiration from the vesicular object by applying negative pressure in the axial vacuum passage. 1. A device for performing micro-operations on a vesicular target , comprising:{'b': 27', '28, 'a. an injection pipette capable of injection or aspiration () such pipette having an opening at its distal end ();'}{'b': 9', '29', '27', '9', '32', '20', '23', '21', '26, 'b. a barrel assembly () having an inside diameter larger than the outside diameter of the pipette so as to create an axial vacuum passage () between the outside of the pipette () and the inside of the barrel assembly (), such axial vacuum passage terminating at a secondary aperture (), the barrel assembly is provided with external O-rings ( and ) situated on either side of a shunt () through the barrel assembly wall, the distal end of the barrel assembly forming an inner tip ();'}{'b': 30', '8', '3', '4', '7, 'c. an outer assembly ...

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Номер записи: 58
26-09-2013 дата публикации

MEDIA FOR CULTURING STEM CELLS

Номер: US20130252329A1
Автор: Amit Michal, Itskovitz-Eldor Joseph
Принадлежит: TECHNION RESEARCH & DEVELOPMENT FOUNDATION LIMITED

Well-defined, xeno-free culture media which comprise a TGF-beta isoform or the chimera formed between IL6 and the soluble IL6 receptor (IL6RIL6), which are capable of mainataining stem cells, and particularly, human embryonic stem cells, in an undifferentiated state are provided. Also provided are cell cultures comprising the culture media and the stem cells and methods of expanding and deriving embryonic stem cells in such well-defined, xeno-free culture media. In addition, the present invention provides methods of differentiating ESCs or EBs formed therefrom for the generation of lineage specific cells. 1. A culture medium comprising an IL6RIL6 chimera and basic fibroblast growth factor (bFGF) , wherein the culture medium is capable of maintaining human pluripotent stem cells in an undifferentiated state.2. A cell culture comprising a human pluripotent stem cell and the culture medium of .3. A method of expanding and maintaining human pluripotent stem cells in an undifferentiated state claim 1 , the method comprising culturing the human pluripotent stem cells in the culture medium of claim 1 , thereby expanding and maintaining the embryonic stem cells in the undifferentiated state.4. The method of claim 3 , wherein said human pluripotent stem cells are human embryonic stem cells.5. A method of deriving a human embryonic stem cell line claim 3 , the method comprising:(a) obtaining a human embryonic stem cell from a pre-implantation stage blastocyst, post-implantation stage blastocyst and/or a genital tissue of a fetus; and{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(b) culturing said human embryonic stem cell in the culture medium of , thereby deriving the embryonic stem cell line.'}6. A method of generating lineage-specific cells from embryonic stem cells claim 3 , the method comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) culturing the embryonic stem cells in the culture medium of , to thereby obtain expanded, undifferentiated embryonic ...

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Номер записи: 59
03-10-2013 дата публикации

Method for Enhancing Umbilical Cord Blood Engraftment

Номер: US20130259825A1
Автор: Tse William
Принадлежит:

A method for enhancing bone marrow cells, peripheral blood, and umbilical cord blood engraftment is disclosed wherein at least one of a granulocyte colony stimulating factor and a granulocyte macrophage colony stimulating factor, and optionally romiplostim, and optionally an erythropoiesis-stimulating agent, are added thereto for enhancing the engraftment of CD44 cells within the bone marrow cells, peripheral blood, and umbilical cord blood. Also provided is a method for enhancing a stem cell infusion by activating an up-regulation of a AF1q/CD44 signaling pathway. A stem cell product is disclosed having treated stem cells conditioned for activating an up-regulation of a AF1q/CD44 signaling pathway. 1. A method for enhancing umbilical cord blood engraftment comprising:providing an amount of an umbilical cord blood;adding to said umbilical cord blood an effective amount of at least one of a granulocyte colony stimulating factor and a granulocyte macrophage colony stimulating factor;optionally adding to said umbilical cord blood an effective amount of romiplostim; andoptionally adding to said umbilical cord blood an effective amount of an erythropoiesis-stimulating agent for forming a treated umbilical cord blood mixture for enhancing the engraftment of CD44 cells within said treated umbilical cord blood mixture.2. The method of including wherein (i) at least one of said granulocyte colony stimulating factor and said granulocyte macrophage colony stimulating factor claim 1 , (ii) said romiplostim claim 1 , and (iii) said erythropoiesis-stimulating agent claim 1 , are added to said umbilical cord blood in any order of addition for forming said treated umbilical cord mixture of said umbilical cord blood claim 1 , at least one of said granulocyte colony stimulating factor and a granulocyte macrophage colony stimulating factor claim 1 , said romiplostim claim 1 , and said erythropoiesis-stimulating agent.3. The method of including wherein (i) at least one of said ...

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Номер записи: 60
03-10-2013 дата публикации

PROGENITOR CELLS FROM WHARTON'S JELLY OF HUMAN UMBILICAL CORD

Номер: US20130259840A1
Автор: Baksh Dolores, DAVIES John E., HOSSEINI Morris, LICKORISH Antony D.S., SARUGASER Rahul
Принадлежит: Tissue Regeneration Therapeutics Inc.

Human progenitor cells are extracted from perivascular tissue of human umbilical cord. The progenitor cell population proliferates rapidly, and harbours osteogenic progenitor cells and MHC−/− progenitor cells, and is useful to grow and repair human tissues including bone. 1. An isolated Wharton's jelly extract , wherein the extract comprises human progenitor cells and is obtained by enzymatic digestion of the perivascular tissue proximal to the vasculature of human umbilical cord.2. The Wharton's jelly extract of claim 1 , wherein the extract is essentially free from cells of umbilical cord blood.3. The Wharton's jelly extract of claim 2 , wherein the extract is obtained by subjecting umbilical cord vasculature bearing proximal Wharton's jelly to enzymatic digestion in a suitable cell extraction medium.4. The Wharton's jelly extract of claim 3 , wherein the step of enzymatic digestion results in the release of cells from the collagen matrix of the Wharton's jelly.5. The Wharton's jelly extract of claim 4 , in which the extraction is performed at 37° C. in phosphate buffered saline and 0.5-10.0 mg/mL collagenase for a period of 1 to 24 hours.6. A method for obtaining one or more human progenitor cells claim 1 , comprising the step of isolating said one or more cells from the Wharton's extract of .7. The method of claim 1 , wherein said method comprises producing a cell population comprising said human progenitor cells comprising the step of culturing at least one said human progenitor cell obtained by the method of .8. A cell population comprising human progenitor cells claim 7 , whenever obtained by the method of .9. An isolated cell population useful as a source of human progenitor cells claim 7 , wherein the cell population is extractable from perivascular tissue of human umbilical cord claim 7 , and has the characteristics of (1) rapid proliferation claim 7 , (2) the presence of osteoprogenitor cells claim 7 , and (3) the presence of immuno-incompetent cells.10. ...

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Номер записи: 61
10-10-2013 дата публикации

FREEZING MEDIUM COMPOSITION FOR CRYOPRESERVING AMNIOTIC FLUID-DERIVED STEM CELLS AND A METHOD FOR CRYOPRESERVING THE SAME

Номер: US20130267008A1
Автор: Shon Yun-Hee, Suh Jang-Soo, Yoo James J.
Принадлежит: KYUNGPOOK NATIONAL UNIVERSITY HOSPITAL

Provided are to a freezing medium composition for cryopreserving amniotic fluid-derived stem cells, which has A lower concentration of MeSO and eliminates fetal bovine serum and at the same time does not induce cryoinjury to fluid-derived stem cells and makes it possible to cryopreserve fluid-derived stem cells for a prolonged time using trehalose, sucrose and catalase, and a method for cryopreserving the same. According to the present invention, AFSCs can be cryopreserved with 1/4 the standard Me2SO concentration with the addition of disaccharides, antioxidants and caspase inhibitors. As a result, the use of Me2SO at low concentrations in cell freezing solutions may support the development of clinical trials of AFSCs. 1. A freezing medium composition for cryopreserving amniotic fluid-derived stem cells , characterized in that it contains trehalose , catalase and zVAD- fmk(Benzyloxycarbonyl-alyl-alanyl-aspartyl-(O-methyl)-fluoromethyl-ketone) with a lower concentration of MeSO.2. The freezing medium composition according to claim 1 , characterized in that the concentration of MeSO is 2.5 to 5% (v/v) and a concentration of trehalose is 60 mmol/L and a concentration of catalase is 100 mg/mL and a concentration of zVAD-fmk is 30 μM.3. A method of cyopreserving amniotic fluid-derived stem cells using the freezing medium composition according to .4. A method of cyopreserving amniotic fluid-derived stem cells using the freezing medium composition according to . This invention relates to a freezing medium composition for cryopreserving amniotic fluid-derived stem cells and a method for cryopreserving the same. More particularly, this invention relates to a freezing medium composition for cryopreserving amniotic fluid-derived stem cells, which has a reduced concentration of MeSO and is free of fetal bovine serum and at the same time does not induce cryoinjury to fluid-derived stem cells, thereby cryopreserving fluid-derived stem cells for a prolonged time using trehalose, ...

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Номер записи: 62
17-10-2013 дата публикации

Culture medium for pluripotent stem cells

Номер: US20130273649A1
Автор: Kim Hoon, Wu Jun, Ying QiLong
Принадлежит: UNIVERSITY OF SOUTHERN CALIFORNIA

The present invention provides culture media and methods of culturing pluripotent stem cells, such as epiblast stem cells (EpiSCs) and embryonic stem cells (ESCs), in order to culture, derive, and reprogram pluripotent stem cells, such as converting ESCs to EpiSCs. 1. A composition , comprising(a) an inhibitor of β-catenin binding to T-cell factors (Tcfs); and(b) a suppressor of glycogen synthase kinase (GSK3) activation.2. The composition of claim 1 , wherein the inhibitor of β-catenin binding to Tcfs is selected from the group consisting of 3 claim 1 ,5 claim 1 ,7 claim 1 ,8-Tetrahydro-2-[4-(trifluoromethyl)phenyl]-4H-thiopyrano[4 claim 1 ,3-d]pyrimidin-4-one (XAV939) claim 1 , 4-(1 claim 1 ,3 claim 1 ,3a claim 1 ,4 claim 1 ,7 claim 1 ,7a-Hexahydro-1 claim 1 ,3-dioxo-4 claim 1 ,7-methano-2H-isoindol-2-yl)-N-8-quinolinyl-Benzamide (IWR-1) claim 1 , and (1R claim 1 ,4r)-4-((2s claim 1 ,3aR claim 1 ,4R claim 1 ,7S claim 1 ,7aS)-1 claim 1 ,3-dioxooctahydro-1H-4 claim 1 ,7-methanoinden-2-yl)-N-(quinolin-8-yl)cyclohexanecarboxamide (53AH) claim 1 , or salts thereof.3. The composition of claim 1 , wherein the suppressor of GSK3 activation is selected from the group consisting of 6-((2-((4-(2 claim 1 ,4-Dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-yl)amino)ethyl)amino)nicotinonitrile (CHIR99021) claim 1 , 2 claim 1 ,6-Pyridinediamine claim 1 , N6-[2-[[4-(2 claim 1 ,4-dichlorophenyl)-5-(1H-imidazol-1-yl)-2-pyrimidinyl]amino]ethyl]-3-nitro- (CHIR 98014) claim 1 , benzyl-2-methyl-1 claim 1 ,2 claim 1 ,4-thiadiazolidine-3 claim 1 ,5-dione (TDZD-8) claim 1 , 3-(2 claim 1 ,4-Dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2 claim 1 ,5-dione (SB216763) claim 1 , 3-[(3-Chloro-4-hydroxyphenyl)amino]-4-(2-nitrophenyl)-1H-pyrrol-2 claim 1 ,5-dione (SB415286) HIR98014 claim 1 , Wnt3a claim 1 , AR-AO144-18 claim 1 , or salts thereof.4. The composition of claim 2 , wherein the suppressor of GSK3 activation is selected from the group consisting of 6-((2-((4-(2 claim 2 ...

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Номер записи: 63
17-10-2013 дата публикации

ADHESION SIGNATURES

Номер: US20130274124A1
Автор: BHATIA Sangeeta N., Malta David Fernandes Braga, Reticker-Flynn Nathan Edward, Schwartz Robert Edward, Underhill Gregory H.
Принадлежит: Massachusetts Institute of Technology

The present invention provides arrays comprising polypeptides associated with extracellular matrix that can be used to isolate, differentiate, or culture certain cell types including stem cells, cancer cells, and/or primary hepatocytes. The array comprises at least a pair of polypeptides that comprise a polypeptide associated with extracellular matrix or functional fragments thereof. The invention also provides for methods of diagnosing and/or prognosing a certain disease or disorder through contacting a cell sample from a patient with an array comprising at least a pair of polypeptides that comprise a polypeptide sequence associated with extracellular matrix or functional fragments thereof. 1. An array of polypeptides , the array comprising: a solid support and a plurality of adhesion sets , wherein each adhesion set comprises two or more different polypeptides comprising a polypeptide sequence associated with the extracellular matrix or a functional fragment thereof , and wherein the adhesion sets are attached to the solid support at an addressable location of the array.2. The array of claim 1 , wherein the solid support is a slide optionally coated with a polymer.3. The array of claim 1 , wherein the polymer is polyacrylamide.4. The array of claim 1 , wherein at least one adhesion set comprises two different polypeptides attached to a solid support.5. The array of claim 1 , wherein the two or more of the different polypeptides comprise at least 10 contiguous amino acids chosen from: collagen I claim 1 , collagen II claim 1 , collagen III claim 1 , collagen IV claim 1 , collagen V claim 1 , collagen VI claim 1 , fibronectin claim 1 , laminin claim 1 , merosin claim 1 , tenascin-R claim 1 , chondroitin sulfate claim 1 , agreccan claim 1 , elastin claim 1 , keratin claim 1 , mucin claim 1 , superfibronectin claim 1 , F-spondin claim 1 , nidogen-2 claim 1 , heparin sulfate claim 1 , biglycan claim 1 , decorin claim 1 , galectin 1 claim 1 , galectin 3 claim 1 , ...

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Номер записи: 64
17-10-2013 дата публикации

Bank of stem cells for producing cells for transplantation having HLA antigens matching those of transplant recipients, and methods for making and using such a stem cell bank

Номер: US20130276154A1
Автор: West Michael
Принадлежит:

Methods for producing stem cell banks, preferably human, which optionally may be transgenic, e.g., comprised of homozygous MHC allele cell lines are provided. These cells are produced preferably from parthenogenic, IVF, or same-species or cross-species nuclear transfer embryos or by dedifferentiation of somatic cells by cytoplasm transfer. Methods for using these stem cell banks for producing stem and differentiated cells for therapy, especially acute therapies, and for screening for drugs for disease treatment are also provided. 1. A stem cell bank comprising a library or plurality of human or non-human animal stem cell lines , each of which is homozygous for at least one MI-IC allele present in a human or non-human animal population , wherein which each member of said plurality of stem cell lines is homozygous for a different set of MHC alleles relative to the remaining members of the plurality of stem cell lines.2. The stem cell bank of which is comprised of a plurality of human stem cell lines each member of which is homozygous for a different combination of MHC alleles than the other members of the library.3. The stem cell bank of which is comprised of at least five different stem cell lines claim 1 , each member homozygous for a different combination of histocompatibility or MHC antigen alleles.4. The human stem cell bank of which is comprised of at least five different stem cell lines claim 2 , each member homozygous for a different combination of histocompatibility or MHC antigen alleles.5. The stem cell bank of which is comprised of at least ten different stem cell lines each member homozygous for a different combination of histocompatibility or MHC antigen alleles.6. The stem cell bank of which is comprised of at least ten different stem cell lines each member homozygous for a different combination of histocompatibility or MHC antigen alleles.7. The stem cell bank of which comprises at least about 100 to 1000 different stem cell lines each homozygous for a ...

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Номер записи: 65
17-10-2013 дата публикации

METHOD AND CULTURE MEDIUM FOR PREPARING MAMMALIAN OVUM OR EMBRYO IN WHICH ZONA PELLUCIDA HAS BEEN THINNED OR ELIMINATED, AND METHOD FOR FERTILIZATION USING MAMMALIAN OVUM PREPARED BY SAME METHOD

Номер: US20130276159A1
Автор: Nakagata Naomi, Takeo Toru
Принадлежит:

Provided are a method for preparing a mammalian ovum or embryo in which zona pellucida has been thinned or eliminated, and a method for fertilization using the mammalian ovum prepared by the aforementioned method. The method for thinning or eliminating zona pellucida of a mammalian ovum or embryo involves treating or culturing the mammalian ovum or embryo (such as an unfertilized ovum, a fertilized ovum, or an embryo in the early stages of development) in a culture medium containing a reducing agent having SH groups (such as reduced glutathione or DTT). The resulting mammalian ovum or embryo (such as an unfertilized ovum, a fertilized ovum, or an embryo in the early stages of development) in which zona pellucida has been thinned or eliminated is capable of realizing an improved fertilization rate and development rate when used for in vitro fertilization, transplantation of a fertilized ovum, or preparation of an embryo in the early stages of development used in the production of a genetically modified animal. 1. A fertilizing method comprising:a. a step of pre-incubating mammal sperms with a culture medium containing 0.1 mM or more and 20 mM or less cyclodextrin and 0.1 mM or more and 10 mM or less calcium,b. a step of adding an unfertilized mammal ovum into a medium containing a reducing agent having SH groups at a concentration of 0.25 mM or more and 500 mM or less in terms of SH equivalent, and preparing an unfertilized ovum treated with the reducing agent, for thinning or elimination of the zona pellucida of the unfertilized ovum, andc. a step of adding the sperms pre-incubated in the step a into the culture medium containing the unfertilized ovum treated with the reducing agent having SH groups in the step b and performing insemination.2. The fertilizing method according to claim 1 , wherein the unfertilized ovum is a mouse unfertilized ovum.3. The fertilizing method according to claim 1 , wherein the cyclodextrin is methyl-β-cyclodextrin.4. The fertilizing ...

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Номер записи: 66
24-10-2013 дата публикации

AGENTS AND METHODS FOR INHIBITING HUMAN PLURIPOTENT STEM CELL GROWTH

Номер: US20130280802A1
Автор: Robins Allan, Schulz Thomas
Принадлежит: ViaCyte, Inc

The present invention relates to compositions and methods for inhibiting or suppressing undifferentiated or pluripotent stem cell growth and proliferation in a differentiated or differentiating cell population or culture. 1. A method for reducing the number or percentage of pluripotent stem cells in a population of cells , comprising:contacting a population of cells comprising at least one pluripotent stem cell with a pluripotent stem cell-selective inhibitor or cytotoxic-agent selected from the group consisting of: caffeic acid, Ivermectin, chelerythrine chloride and combinations thereof, thereby reducing the number or percentage of pluripotent stem cells in the population of cells.2. The method of claim 1 , wherein the pluripotent stem cell is a primate cell.3. The method of claim 2 , wherein the primate is a human.4. The method of claim any of claim 1 , wherein the pluripotent stem cell is an ES cell or an iPS cell.56-. (canceled)7. The method of claim 1 , wherein the population of cells is incubated in a cell culture medium containing at least about 1 μM caffeic acid claim 1 , at least about 1 μM Ivermectin or at least 5 about μM chelerythrine chloride.89-. (canceled)10. The method of claim 1 , wherein the population of cells further comprises differentiating or differentiated cells.11. The method of claim 10 , wherein the differentiating or differentiated cells are derived from the pluripotent stem cells.12. The method of claim 10 , wherein the pluripotent stem cell-selective inhibitor or cytotoxic agent has no effect on the viability or proliferation of the differentiating or differentiated cells.1314-. (canceled)15. The method of claim 10 , wherein the pluripotent stem cell-selective inhibitor or cytotoxic agent does not inhibit the differentiation or differentiation potential of the cells in the population.1620-. (canceled)20. The method of claim 10 , wherein the differentiating or differentiated cells comprise at least one cell type selected from: ...

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Номер записи: 67
24-10-2013 дата публикации

DEFINED CELL CULTURING SURFACES AND METHODS OF USE

Номер: US20130280803A1
Автор: ABRAHAM ELIZABETH, QIAN SUSAN XIUQI, Sanyal Supama, SAXENA DEEPA
Принадлежит:

In one aspect, there is provided a cell culturing substrate including: 1. A cell culturing substrate comprising:a cell culture surface having a film attached thereto, wherein said film comprises one or more plasma polymerized monomers; anda coating on said film-coated surface, said coating deposited from a coating solution comprising one or more extracellular matrix proteins and an aqueous solvent, wherein the total extracellular matrix protein concentration in the coating solution is about 1 ng/mL to about 1 mg/mL.2. The cell culturing substrate of claim 1 , wherein said one or more monomers is selected from the group consisting of acrylic acid claim 1 , methacrylic acid claim 1 , acetic acid claim 1 , vinylacetic acid and combinations thereof.3. The cell culturing substrate of claim 1 , wherein said one or more monomers is selected from the group consisting of allylamine claim 1 , methylamine claim 1 , propylamine claim 1 , heptylamine and diaminopropane.4. The cell culturing substrate of claim 1 , wherein said monomer is selected from the group consisting of alkanes claim 1 , alkenes claim 1 , dienes claim 1 , styrenes and combinations thereof.5. The cell culturing substrate of claim 1 , wherein said one or more monomers is selected from the group consisting of amines claim 1 , hydrocarbons and combinations thereof claim 1 , wherein the ratio of amine to hydrocarbon is between about 100:0 and about 20:80.6. The cell culturing substrate of claim 1 , wherein the total protein concentration in the coating composition is about 1 μg/mL to about 300 μg/mL.7. The cell culturing substrate of claim 1 , wherein the total protein concentration in the coating composition is about 5 μg/mL to about 200 μg/mL.8. The cell culturing substrate of claim 1 , wherein the coating composition comprises extracellular matrix proteins selected from the group consisting of natural claim 1 , recombinant claim 1 , synthetic extracellular matrix proteins and combinations thereof.9. The cell ...

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Номер записи: 68
24-10-2013 дата публикации

COMPOSITION FOR REPROGRAMMING SOMATIC CELLS TO GENERATE INDUCED PLURIPOTENT STEM CELLS, COMPRISING Oct4 IN COMBINATION WITH Bmi1 OR ITS UPSTREAM REGULATOR, AND METHOD FOR GENERATING INDUCED PLURIPOTENT STEM CELLS USING THE SAME

Номер: US20130280808A1
Автор: Heo June Seok, Jun Eun Kyoung, Kim Jun Sung, LEE Jung Han, Moon Jai-Hee, Yoon Byung Sun, You Seungkwon
Принадлежит:

Disclosed is a composition for reprogramming somatic cells to generate embryonic stem cell-like cells, comprising: a) a Bmi1 (B cell-specific Moloney murine leukemia virus integration site 1) protein or a nucleic acid molecule coding for Bmi1; and b) an Oct4 protein or a nucleic acid molecule coding for Oct4. Also, a method is provided for reprogramming somatic cells to generate embryonic stem cell-like cells using the composition. In addition to reducing the number of the genetic factors conventionally needed, the composition and method allow the generation of pluripotent embryonic stem cell-like cells which have high potential in the cell therapy of various diseases. 13-. (canceled)4. A method for reprogramming somatic cells to generate embryonic stem cell-like cells by introducing a Bmi1 (B cell-specific Moloney murine leukemia virus integration site 1) gene and an Oct4 gene into the somatic cells.5. The method according to claim 4 , comprising:(i) culturing fibroblasts in a medium(ii) infecting the fibroblasts with a packaging cell into which respective vectors carrying a Bmi1 gene and an Oct4 gene are transfected; and(iii) culturing the infected fibroblasts under the conditions used to culture embryonic stem cells.6. The method according to claim 5 , wherein the vector carrying a Bmi1 gene is constructed by inserting a nucleic acid molecule coding for a Bmi1 protein into a pBabe puro vector and the vector carrying an Oct4 gene is constructed by inserting a nucleic acid molecule coding for an Oct4 protein into a pBabe neo vector.7. The method according to claim 5 , wherein the packaging cell is a PT67 packaging cell.89-. (canceled)10. A method for reprogramming somatic cells to generate neural stem cells claim 5 , comprising:introducing a Bmi1 (B cell-specific Moloney murine leukemia virus integration site 1) gene into the somatic cells; andculturing the somatic cells in a condition used to culture neural stem cells.1113-. (canceled)14. A method for ...

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Номер записи: 69
31-10-2013 дата публикации

METHOD FOR DIFFERENTIATING HUMAN EMBRYONIC STEM CELLS INTO ß-CELLS FOR THE TREATMENT OF TYPE I DIABETES

Номер: US20130287743A1
Автор: Colton Clark K., Dilenno Amanda, Millman Jeffrey R.
Принадлежит: Massachusetts Institute of Technology

The invention provides inter alia methods for differentiating embryonic stem cells into insulin producing cells, as well as compositions comprising such cells, and therapeutic uses of such compositions. 1. A method comprising{'sub': '2', 'differentiating embryonic stem cells to posterior foregut endoderm in vitro, at low oxygen partial pressure (pO), and'}{'sub': '2', 'differentiating posterior foregut endoderm to insulin+ and/or C-peptide+ cells in vitro, at higher pO.'}2. The method of claim 1 , wherein the embryonic stem cells are differentiated on an oxygen permeable membrane.3. The method of claim 2 , wherein the oxygen permeable membrane is an oxygen permeable silicone rubber membrane.4. The method of claim 2 , wherein the oxygen permeable silicone rubber membrane is coated with extracellular matrix (ECM).5. The method of claim 1 , wherein the embryonic stem cells are human embryonic stem cells.6. A method comprising{'sub': '2', 'differentiating human embryonic stem cells to a definitive endoderm stage at low oxygen partial pressure (pO).'}7. The method of claim 6 , further comprising differentiating the definitive endoderm stage to a primitive gut tube stage at low oxygen partial pressure (pO).8. The method of claim 7 , further comprising differentiating the primitive gut tube stage to a posterior foregut stage at low oxygen partial pressure (pO).9. The method of claim 8 , further comprising differentiating the posterior foregut stage to a pancreatic endoderm stage at an increased oxygen partial pressure (pO).10. The method of claim 9 , further comprising differentiating the pancreatic endoderm stage to an insulin+ and/or C-peptide+ cell stage at an increased oxygen partial pressure (pO).11. The method of claim 6 , wherein the low oxygen partial pressure (pO) is 36 mmHg.12. The method of claim 7 , wherein the low oxygen partial pressure (pO) is 36 mmHg.13. The method of claim 8 , wherein the low oxygen partial pressure (pO) is 36 mmHg.14. The method of claim ...

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Номер записи: 70
31-10-2013 дата публикации

De Novo Anembryonic Trophoblast Vesicles and Methods of Making and Using Them

Номер: US20130287822A1
Автор: Robins Jared
Принадлежит:

Anembryonic, de novo, trophoblast vesicles are further characterized by (a) having the functional capacity for implantation, (b) being composed of a substantially perfect sphere having a hollow, acellular center, (c) having a cellular rim containing viable cells that are proliferating, and (d) having numerous tight-junctions among the viable cells in the rim. Embryos can be made by seeding trophoblast cells in a non-adherent cell aggregation device; incubating the cells in the device until the cells form functional anembryonic de novo trophoblast vesicles; and injecting an inner cell mass or embryonic stem cells into the functional anembryonic de novo trophoblast vesicles to thereby make the embryos. 1. Anembryonic , de novo , trophoblast vesicles further characterized by (a) having the functional capacity for implantation , (b) being composed of a substantially perfect sphere having a hollow , acellular center , (c) having a cellular rim containing viable cells that are proliferating , and (d) having numerous tight-junctions among the viable cells in the rim.2. Anembryonic claim 1 , de novo claim 1 , trophoblast vesicles according to wherein the functional capacity for implantation is exhibited by production of MMP-2 and MMP-9.3. Anembryonic claim 1 , de novo claim 1 , trophoblast vesicles according to wherein the vesicles demonstrate Na-K ATPase activity.4. Anembryonic claim 1 , de novo claim 1 , trophoblast vesicles according to wherein the vesicles implant when transferred into a uterine horn of a pseudo-pregnant claim 1 , immunodeficient mouse surrogate.5. A method of making embryos comprising the steps of:(a) seeding trophoblast cells in a non-adherent cell aggregation device;(b) incubating the cells in the device until the cells form functional anembryonic de novo trophoblast vesicles; and(c) injecting an inner cell mass or embryonic stem cells into the functional anembryonic de novo trophoblast vesicles to thereby make the embryos.6. The method of wherein ...

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Номер записи: 71
31-10-2013 дата публикации

FUSION PROTEIN AND USE THEREOF

Номер: US20130288306A1
Автор: Bosnali Manal, Edenhofer Frank, Peitz Michael, Thier Marc-Christian
Принадлежит:

The invention relates to a fusion protein and a method for the generation of the fusion protein of the invention. Further, the invention relates to the use of the fusion protein of the invention for the generation of induced pluripotent cells. Moreover, the invention relates to a composition comprising at least one fusion protein of the invention. 1. A Fusion protein comprising:a) a protein transduction domain (PTD) comprising 6 to 12 basic amino acids; andb) at least one transcription factor.2. The fusion protein of claim 1 , wherein the PTD further comprises hydrophobic amino acids.3. The fusion protein of claim 1 , wherein the PTD further comprises at least one of TAT claim 1 , Penetratin claim 1 , HSV-VP22 claim 1 , Transportan claim 1 , K-FGF claim 1 , Oligoarginine claim 1 , Arg-9 claim 1 , and peptides consisting of combinations of Arginine and Lysine residues.4. The fusion protein of claim 1 , wherein the PTD is comprised of no more than 20 amino acids.5. The fusion protein of claim 1 , wherein the transcription factor is selected from the group consisting of: stem cell factors claim 1 , such as Oct4 claim 1 , Sox2 claim 1 , nanog claim 1 , klf4 claim 1 , lin28 claim 1 , hTERT claim 1 , myc claim 1 , SV40; differentiation factors claim 1 , such as hox-genes claim 1 , pax genes; reprogramming factors claim 1 , such as HMGB claim 1 , Msx1; tumor suppressors claim 1 , such as p53 claim 1 , Pten; and anti-apoptose factors claim 1 , such as bcl-2.6. The fusion protein of claim 5 , wherein the transcription factor is one of Sox2 and Oct4.7. The fusion protein of claim 1 , further comprising a nuclear localization signal (NLS).8. The fusion protein of claim 1 , wherein the fusion protein comprises an artificial trans activation domain (ATAD).9. The fusion protein of claim 1 , further comprising a linker selected from the group consisting of Proline claim 1 , Glycine claim 1 , Alanine claim 1 , Leucine claim 1 , Glutamic acid claim 1 , Threonine claim 1 , and Serine ...

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Номер записи: 72
31-10-2013 дата публикации

PROTEIN DELIVERY SYSTEM TO GENERATE INDUCED PLURIPOTENT STEM (iPS) CELLS OR TISSUE-SPECIFIC CELLS

Номер: US20130288374A1
Автор: Chang Yung-Nien, Oyler George A.
Принадлежит: Synaptic Research, LLC

A novel protein delivery system to generate induced pluripotent stem (iPS) cells is described. The delivery system comprises a construct with a receptor binding domain that recognizes a receptor in a somatic cell, a translocation domain that allows the transfer of an inducer into the cytosolic space, and a cargo bearing domain to which the inducer is attached and facilitates transfer of the inducer into the cell. 1. A method for generating induced Pluripotent Stem (iPS) cells , comprising:{'i': 'Clostridium', 'contacting somatic cell with a construct that comprises a receptor binding domain, a translocation domain, a cargo bearing domain, and an inducer, wherein the receptor binding domain, translocation domain, and cargo domain are atoxic toxin domains.'}2. The method of claim 1 , further comprising the step of assessing the pluripotency of the somatic cells after contact with the construct.3. The method of claim 1 , wherein the construct is transfected into the somatic cell during contact.4ClostridiumC. difficile. The method of claim 1 , wherein the toxin is a toxin.5C. difficile. The method of claim 4 , wherein the toxin is TcdA.6C. difficile. The method of claim 4 , wherein the toxin is TcdB.7. The method of claim 1 , wherein said cargo bearing domain is an inactive exotoxin domain.8ClostridiumC. botulinum. The method of claim 1 , wherein the toxin is a toxin.9C. botulinum. The method of claim 8 , wherein the toxin is selected from the group consisting of BoNTA claim 8 , BoNTB claim 8 , BoNTC claim 8 , BoNTD claim 8 , BoNTE claim 8 , BoNTF and BoNTG.10C. botulinum. The construct of claim 8 , wherein a receptor binding domain of the toxin is replaced with a non-specific receptor binding domain.11Clostridium. The method of claim 1 , wherein the toxin is a clostridial C2 toxin.12. The method of claim 1 , wherein the inducer is Oct3/4.13. The method of claim 12 , wherein the construct has a sequence comprising the amino acid sequence of SEQ ID No: 1.14. The method ...

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Номер записи: 73
07-11-2013 дата публикации

SERUM-FREE MAMMALIAN CELL CULTURE MEDIUM, AND USES THEREOF

Номер: US20130295667A1
Автор: Dzimian Joyce, Epstein David, FIKE Richard, GODWIN GLENN, Gorfien Stephen, Gruber Dale, PRICE Paul
Принадлежит:

The present invention provides a cell culture medium formulation that supports the in vitro cultivation, particularly in suspension, of mammalian cells, particularly epithelial cells and fibroblast cells, and methods for cultivating mammalian cells in suspension in vitro using these media. The media comprise a basal medium and a polyanionic or polyanionic compound, preferably a polysulfonated or polysulfated compound, and more preferably dextran sulfate. The present invention also provides chemically defined, protein-free eukaryotic cell culture media comprising an iron chelate and zinc, which is capable of supporting the growth (and particularly the high-density growth of mammalian cells) in suspension culture, increasing the level of expression of recombinant protein in cultured cells, and/or increasing virus production in cultured cells. 137-. (canceled)38. A kit for the cultivation of a CHO cell in suspension in vitro , said kit comprising one or more containers , wherein a first container contains a serum-free and protein-free cell culture medium comprising a transferrin substitute and an insulin substitute wherein said medium supports the cultivation of said CHO cell in suspension without supplementation of serum and without supplementation of protein , wherein said transferrin substitute comprises iron or an iron containing compound and said insulin substitute comprises zinc or a zinc containing compound , and wherein the medium supports increased CHO cell growth in suspension as compared to culture in FMX-8 medium , and wherein the cultivation of the CHO cell does not require that the CHO cell express an exogenous recombinant protein.39. (canceled)40. The kit of claim 38 , further comprising a second container containing at least one CHO cell.41. The kit of claim 38 , further comprising a second container containing at least one polyanionic or polycationic compound.42. The kit of claim 38 , wherein said increased CHO cell growth is to about 1.5×10to 2× ...

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Номер записи: 74
07-11-2013 дата публикации

SYSTEMS AND METHODS FOR PROCESSING CELLS

Номер: US20130295673A1
Автор: Meade John, Taghizadeh Rouzbeh R.
Принадлежит:

The present invention efficiently and cost-effectively extracts and collects cells from a tissue. The inventors have discovered that the tissue can be effectively fragmented and the resulting cells can be purified using a system or kit with multiple components. An advantage of the present invention is that tissue processing takes place in a closed system such that sterility can be maintained throughout the process, even if certain components are removed during processing, for example through the use of valves, clamps, and heat seals. Furthermore, any or all of the steps can be automated or manually accomplished, according to the specific needs of the application or the user. 1. A tissue mincing tool comprising:a compartment for a tissue sample;a cutting surface at one end of the compartment; anda sterile, sealed container,wherein the cutting surface separates the compartment from the sterile, sealed container, such that a tissue sample that passes through the cutting surface is deposited within the sterile, sealed container.2. The tissue mincing tool of claim 1 , wherein the cutting surface is dimensioned to mince the tissue sample into fragments having an average cross-section no greater than four square millimeters.3. The tissue mincing tool of claim 1 , wherein the cutting surface is dimensioned to mince the tissue sample into fragments claim 1 , and further comprising a second cutting surface for reducing an average cross-section of the fragments.4. The tissue mincing tool of claim 1 , further comprising at least one mincer screen positioned in proximity to the cutting surface.5. The tissue mincing tool of claim 4 , wherein the at least one mincer screen is moveable towards the tissue sample.6. The tissue mincing tool of claim 1 , further comprising a suction cup for stabilizing the tissue mincing tool during operation thereof.7. The tissue mincing tool of claim 1 , wherein the sterile claim 1 , sealed container comprises at least one sealed access port ...

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Номер записи: 75
14-11-2013 дата публикации

Compositions and methods for growing human embryonic cells

Номер: US20130302887A1
Автор: Michael Cohen, Michael Shamblott
Принадлежит: Individual

Methods for deriving and cultivating human embryonic stem (ES) cells and maintaining their pluripotency in culture is provided by utilizing secreted products obtained from the culture medium of human embryonic gem (EG) cell derivatives, such as embryoid body-derived cells. Substrates include compounds such as collagen I, fibronectin, or superfibronectin, or extracellular matrix, typically human derived.

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Номер записи: 76
14-11-2013 дата публикации

Stem cells & matrix from cord tissue

Номер: US20130302890A1
Автор: Raymond MOUZANNAR
Принадлежит: Stem Cell Reserve LP LLC

Methods for isolating and culturing stem cells, making tissue matrix, making matrix infused with stem cells, and methods of stem cell therapy are provided.

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Номер записи: 77
21-11-2013 дата публикации

GYNOGENETIC OR ANDROGENETIC PRODUCTION OF PLURIPOTENT CELLS AND CELL LINES, AND USE THEREOF TO PRODUCE DIFFERENTIATED CELLS AND TISSUES

Номер: US20130309206A1
Автор: Burnside Amy, Cibelli Jose, Robl James M.
Принадлежит: University of Massachusetts

Methods for obtaining pluripotent (embryonic stem) cells from parthenogenetic embryos, especially primates, are provided. These cells are useful for producing differentiated cells, tissues and organs, especially human and non-human primate cells, tissues and organs. 1. A method for producing pluripotent (ES) cells that can be used to produce differentiated cells and tissues comprising:(a) obtaining a haploid cell in metaphase II that comprises DNA derived from a single individual male or female, which optionally may be genetically modified;(b) activating said haploid cell by a method selected from the group consisting of (1) conditions that do not result in second polar body extrusion; (2) conditions that provide for polar body extrusion but in the presence of an agent that inhibits polar body extrusion, and (3) conditions that prevent the initial cleavage, and culturing said activated cell to produce a gynogenetic or androgenetic embryo comprising a discernible trophectoderm and an inner cell mass;(c) isolating said inner cell mass or cells therefrom and transferring said inner cell mass or cells to an in vitro media that inhibits differentiation of said inner cell mass derived therefrom; and(d) culturing said inner cell mass cells or cells derived therefrom to maintain said cells in an undifferentiated pluripotent state.2. The method of claim 1 , wherein the metaphase II cell is an oocyte or blastomere.3. The method of claim 2 , wherein the haploid cell is a human claim 2 , non-human primate claim 2 , bovine claim 2 , porcine claim 2 , or ovine oocyte or blastomere.4. The method of claim 3 , wherein the haploid DNA derived from a single individual is human claim 3 , bovine claim 3 , primate claim 3 , ovine claim 3 , or porcine.5. The method of claim 4 , wherein the cell is a human or bovine oocyte and the haploid DNA is human DNA.6. The method of claim 1 , where said activation conditions include the use of DMAP (phosphorylation inhibitor) or other compound that ...

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Номер записи: 78
21-11-2013 дата публикации

METHOD FOR CULTURING HUMAN PLURIPOTENT STEM CELLS

Номер: US20130309768A1
Автор: Furue Miho, Kinehara Masaki
Принадлежит: National Institute of Biomedical Innovation

An aim of the present invention is to provide a method for culturing a human pluripotent stem cell while maintaining an undifferentiated state, more efficiently than conventional methods, and a kit therefor. The pluripotency of a stem cell was found to be maintained at a high rate, regardless of experimenter's proficiency in culturing techniques, by (a) culturing a human pluripotent stem cell in a first medium which a medium for pluripotent stem cell comprising activin; (b) replacing the first medium with a second medium which is a medium for pluripotent stem cell comprising no activin, and culturing the human pluripotent stem cell, (c) subculturing the human pluripotent stem cell into the first medium, and then repeating the above (b) and (c) sequentially. 1. A method for culturing a human pluripotent stem cell while maintaining an undifferentiated state , comprising the steps (a) to (c) , wherein after the step (a) , the steps (b) and (c) are sequentially repeated:(a) a step of culturing a human pluripotent stem cell in a first medium comprising a basal medium for pluripotent stem cell supplemented with an activin-containing supplement;(b) a step of replacing the first medium with a second medium comprising the basal medium for pluripotent stem cell supplemented with an activin-free supplement, and culturing the human pluripotent stem cell; and(c) a step of subculturing the human pluripotent stem cell into the first medium.2. The method for culturing according to claim 1 , wherein the supplements comprise fibronectin claim 1 , insulin claim 1 , transferrin claim 1 , 2-mercaptoethanol claim 1 , 2-ethanolamine claim 1 , sodium selenite and an albumin-conjugated oleic acid.3. The method according to claim 2 , wherein fibronectin is used without being coated on an inside of a culture vessel.4. The method according to claim 2 , wherein the supplements further comprise a protein kinase C inhibitor.5. The method for culturing according to claim 1 , wherein the basal ...

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Номер записи: 79
28-11-2013 дата публикации

AUTOMATED METHOD AND APPARATUS FOR EMBRYONIC STEM CELL CULTURE

Номер: US20130316445A1
Автор: BEARDSLEY Nathaniel, BERGENDAHL Veit, DAIGH Christine, FITZGERALD Megan
Принадлежит: CELLULAR DYNAMICS INTERNATIONAL, INC.

The invention concerns methods for automated culture of embryonic stem cells (ESCs) such as human ESCs. In some aspects, methods of the invention employ optimized culture media and limited proteolytic treatment of cells to separate cell clusters for expansion. Automated systems for passage and expansion of ESCs are also provided. 139-. (canceled)40. An apparatus for automated maintenance and expansion of pluripotent cells comprising:a) an incubator;b) a liquid handler unit;c) one or more reservoirs, said reservoirs comprising i) a viable population of pluripotent cells, ii) a defined media, the defined media being one in which pluripotent cells can be cultured and maintained in an undifferentiated state, and iii) one or more of a protease, protease inhibitor and a Rho-associated kinase (ROCK) inhibitor; andd) a controller in communication with the liquid handler unit, wherein the controller is configured to direct the liquid handler unit to effect the automated expansion and maintenance of the pluripotent cells.41. The apparatus of claim 40 , wherein the controller comprises a computer-readable medium including an operating program.42. The apparatus of claim 40 , wherein the controller directs at least one of: the liquid handler unit claim 40 , the one or more reservoirs claim 40 , and the incubator to i) remove media from a pluripotent cell population; ii) contact pluripotent cells with a proteolytic enzyme; and iii) incubate pluripotent cells with a proteolytic enzyme to separate cell clusters.43. The apparatus of claim 40 , wherein the apparatus further comprises a mechanical agitator and aspirator claim 40 , and the controller further directs at least one of: the liquid handler unit claim 40 , the one or more reservoirs claim 40 , and the incubator to subject the pluripotent cells to mechanical agitation or aspiration to further separate cell clusters.44. The apparatus of claim 40 , wherein the controller further directs at least one of: the liquid handler unit ...

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Номер записи: 80
05-12-2013 дата публикации

METHOD OF GENERATING INDUCED PLURIPOTENT STEM CELLS AND DIFFERENTIATED CELLS

Номер: US20130323782A1
Автор: Esteban Miguel, Grillari Johannes, Grillari Regina, Pei Duanqing, Zhou Ting
Принадлежит:

Methods for generating iPSCs and differentiated cells of interest by reprogramming donor cells that have been obtained in a non-invasive manner. In particular, the donor cells are exfoliated epithelial urine cells. The differentiated cells can be obtained by differentiation of the reprogrammed iPSCs or by direct reprogramming the urine cells. 1. A method for generating differentiated cells of interest from donor cells of another cell type , comprising:i) expanding cells from a cell source obtained from a donor in a non-invasive manner, wherein said cell source is selected from the group consisting of urine, feces, saliva, hair, nasal secretion, cerumen, lacrimal fluid and the vaginal tract, and a) reprogramming said expanded cells to become induced pluripotent stem cells (iPSCs) and differentiating said iPSCs into said cells of interest, or by', 'b) directly reprogramming said cells to become differentiated cells of interest., 'ii) generating differentiated cells from said expanded cells by2. The method of claim 1 , wherein said donor cells of step i) are exfoliated cells derived from urine.3. The method of claim 2 , wherein said cells are selected from the group consisting of epithelial cells claim 2 , fibroblastoid cells claim 2 , and cancer cells.4. The method of claim 1 , wherein said reprogramming is effected by a mixture of exogenous reprogramming factors comprising factors selected from the group consisting of an Oct family gene claim 1 , a Klf family gene claim 1 , and a Sox family gene.5. The method of claim 1 , wherein said reprogramming is effected by a mixture of exogenous reprogramming factors comprising factors selected from the group consisting of Sox2 claim 1 , Oct4 claim 1 , Klf4 claim 1 , c-Myc and Nanog.6. The method of claim 4 , wherein said reprogramming involves expression of reprogramming factors by the cell without genomic integration and/or without expression of oncogenes.7. The method of claim 4 , wherein said reprogramming additionally ...

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Номер записи: 81
05-12-2013 дата публикации

REPROGRAMMING CELLS

Номер: US20130323833A1
Автор: Ding Sheng, Zhu Saiyong
Принадлежит:

The present invention provides for methods, compositions, and kits for producing an induced pluripotent stem cell from a non-pluripotent mammalian cell using a 3′-phosphoinositide-dependent kinase-1 (PDK1) activator or a compound that promotes glycolytic metabolism as well as other small molecules. 1. A method of inducing a non-pluripotent mammalian cell into an induced pluripotent stem cell , the method comprising:(a) contacting the non-pluripotent cell with an Oct polypeptide or introducing into the non-pluripotent cell a polynucleotide encoding an Oct polypeptide; and(b) contacting the non-pluripotent cell with a 3′-phosphoinositide-dependent kinase-1 (PDK1) activator under conditions sufficient to induce the cell to become a pluripotent stem cell, thereby inducing the non-pluripotent mammalian cell into an induced pluripotent stem cell.2. The method of claim 1 , wherein the PDK1 activator is:(a) an allosteric PDK1 activator; or(b) (Z)-5-(4-Chlorophenyl)-3-phenylpent-2-enoic acid) (“PS48”), (Z)-5-(4-Bromo-2-fluorophenyl)-3-phenylpent-2-enoic acid (“PS08”), 2-(3-(4-Chlorophenyl)-3-oxo-1-phenylpyropylthio)acetic acid, (Z)-5-(Napthalen-2-yl)-3-phenylpent-2-enoic acid (“12Z”), or (Z)-5-(1H-Indol-3-yl)-3-phenylpent-2-enoic acid (“13Z”).34-. (canceled)5. The method of claim 1 , further comprising contacting the non-pluripotent cell with (a) a TGFβ receptor/ALK5 inhibitor or a MEK inhibitor claim 1 , or (b) a TGFβ receptor/ALK5 inhibitor and a MEK inhibitor.68-. (canceled)9. The method of claim 1 , further comprising contacting the non-pluripotent cell with a histone deacetylase (HDAC) inhibitor.1014-. (canceled)15. The method of claim 1 , further comprising one or more of:(a) contacting the non-pluripotent cell with a Klf polypeptide or introducing into the non-pluripotent cell a polynucleotide encoding a Klf polypeptide;(b) contacting the non-pluripotent cell with a Sox polypeptide or introducing into the non-pluripotent cell a polynucleotide encoding a Sox ...

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Номер записи: 82
19-12-2013 дата публикации

Cell Culture

Номер: US20130337529A1
Автор: Choo Yen
Принадлежит:

A method for determining the effect of a plurality of culture conditions on a cell, comprising the steps of: a) providing a first set of groups of cell units each comprising one or more cells, and exposing said groups to desired culture conditions; (b) pooling two or more of said groups to form at least one second pool; (c) subdividing the second pool to create a further set of groups of cell units; (d) exposing said further groups to desired culture conditions; (e) optionally, repeating steps (b)-(d) iteratively as required; and (f) optionally assessing the effect on a given cell unit of the culture conditions to which it has been exposed. 146.-. (canceled)47. A method for obtaining differentiated cells from stem cells or progenitor cells in vitro , comprising the steps of:(a) growing stem cells or progenitor cells in a culture medium adhered to a microcarrier or a bead, confined within a medium permeable barrier or entrapped;(b) transferring the cells in step (a) from one culture medium to another;(c) repeating step (b); and(d) obtaining the differentiated cells.48. A method according to claim 47 , wherein the differentiated cells are isolated by enzymatic detachment from the microcarrier or bead.49. A method according to claim 47 , wherein the differentiated cells are isolated by digestion of the microcarrier.50. A method according to claim 48 , wherein the differentiated cells are isolated by digestion of the microcarrier.51. The method according to claim 47 , wherein the microcarrier comprises a plastic claim 47 , a polymer claim 47 , a macromolecule such as cellulose claim 47 , dextran claim 47 , agarose claim 47 , or acrylamide claim 47 , or inorganic materials such as glass.52. The method according to claim 47 , wherein the cells are entrapped in cellulose fibres such as DEAE claim 47 , TLC claim 47 , QAE claim 47 , TEAE or in ceramic cartridges.53. The method according to claim 47 , wherein the cells are bipotent claim 47 , pluripotent or totipotent cells. ...

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Номер записи: 83
19-12-2013 дата публикации

ADHERENT CELLS FROM PLACENTA TISSUE AND USE THEREOF IN THERAPY

Номер: US20130337558A1
Автор: Drori-Carmi Nirit, Meiron Moran, Ofir Rachel, Toren Amir
Принадлежит: PLURISTEM LTD.

A method of culturing adherent cells from a placenta or adipose tissue is disclosed. The method comprises culturing the adherent cells from the placenta or adipose tissue under 3 dimensional (3D) culturing conditions which allow cell expansion, the conditions comprising perfusion. 1. A population of cells made b the method comprising culturing adherent cells from placenta or adipose tissue under three-dimensional (3D) culturing conditions which allow for cell expansion , said conditions comprising perfusion , wherein the rate of said perfusion is adjusted according to the glucose concentration of the culture medium.221-. (canceled) This application claims the benefit of priority from Provisional Patent Application 61/202,050 filed Jan. 23, 2009 and Provisional Patent Application 61/136,375 filed Sep. 2, 2008.The contents of all of the above documents are incorporated by reference as if fully set forth herein.The present invention, in some embodiments thereof, relates to adherent cells of placenta tissue and, more particularly, but not exclusively, to methods of culturing same and using same for treatment.In recent years, considerable activity has focused on the therapeutic potential of mesenchymal stromal cells (MSCs) for various medical applications including tissue repair of damaged organs such as the brain, heart, bone and liver and in support of bone marrow transplantations (BMT). MSCs, a heterogeneous population of cells obtained from e.g. bone marrow, adipose tissue, placenta, and blood, is capable of differentiating into different types of cells (e.g. reticular endothelial cells, fibroblasts, adipocytes, osteogenic precursor cells) depending upon influences from various bioactive factors. Accordingly, MSCs have been widely studied in regenerative medicine as the foundation to build new tissues such as bone, cartilage and fat for the repair of injury or replacement of pathologic tissues and as treatment for genetic and acquired diseases [Fibbe and Noort, Ann N ...

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Номер записи: 84
19-12-2013 дата публикации

METHODS FOR CULTURING UNDIFFERENTIATED CELLS USING SUSTAINED RELEASE COMPOSITIONS

Номер: US20130337562A1
Автор: Stern Jeffrey, Stern Sally Temple
Принадлежит: REGENERATIVE RESEARCH FOUNDATION

Methods for culturing undifferentiated mammalian cells, such as stem and progenitor cells, are provided. The methods involve incubating the cell in the presence of a sustained release composition containing at least one growth factor, wherein the sustained release composition continuously releases the growth factor(s), and wherein the presence of the sustained level of growth factor maintains the cell in an undifferentiated state. 136-. (canceled)37. A method for culturing a stem or progenitor cell , which method comprises:(a) adding, to a culture of stem or progenitor cells, a sustained release composition comprising at least one growth factor that maintains the cells in an undifferentiated state; and(b) repeating the step of adding the sustained release composition such that the cells are maintained in an undifferentiated state.38. The method of claim 37 , wherein the sustained release composition is added to the culture at least once a week.39. The method of claim 37 , wherein the sustained released composition is added to the culture not more often than every other day.40. The method of claim 37 , wherein the sustained release composition is added to the culture not more often than every three days.41. The method of claim 37 , wherein the sustained release composition comprises at least one growth factor selected from the group consisting of: epidermal growth factor (EGF) claim 37 , platelet-derived growth factor (PDGF) claim 37 , sonic hedgehog (Shh) claim 37 , leukemia inhibitor factor (LIF) claim 37 , Wnt protein claim 37 , and fibroblast growth factor 2 (FGF2).42. The method of claim 37 , wherein the sustained release compositions comprises at least fibroblast growth factor 2 (FGF2).43. The method of claim 42 , wherein the sustained release composition further comprises one or more additional growth factors.44. The method of claim 43 , wherein the one or more additional growth factors include one or more growth factors selected from the group consisting of: ...

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Номер записи: 85
19-12-2013 дата публикации

ISOLATION, CULTIVATION AND USES OF STEM/PROGENITOR CELLS

Номер: US20130337563A1
Автор: LIM Ivor Jiun, PHAN Toan-Thang
Принадлежит:

The present invention relates to a method for isolating stem/progenitor cells from the amniotic membrane of umbilical cord, wherein the method comprises separating the amniotic membrane from the other components of the umbilical cord in vitro, culturing the amniotic membrane tissue under conditions allowing cell proliferation, and isolating the stem/progenitor cells from the tissue cultures. The isolated stem cell cells can have embryonic stem cell-like properties and can be used for various therapeutic purposes. In one embodiment, the invention relates to the isolation and cultivation of stem cells such as epithelial and/or mesenchymal stem/progenitor cells under conditions allowing the cells to undergo mitotic expansion. Furthermore, the invention is directed to a method for the differentiation of the isolated stem/progenitor cells into epithelial and/or mesenchymal cells. 1. A method for isolating stem/progenitor cells from the amniotic membrane of umbilical cord , the method comprising:(a) separating the amniotic membrane from the other components of the umbilical cord in vitro;(b) culturing the amniotic membrane tissue obtained in step (a) under conditions allowing cell proliferation; and(c) isolating the stem/progenitor cells.2. The method of claim 1 , further comprising:(a″) separating the cells of the amniotic membrane tissue before cultivation by a method selected from the group consisting of enzymatic separation and direct tissue explant.3. The method of claim 1 , wherein the stem/progenitor cells have embryonic stem cell-like properties.4. The method of claim 1 , wherein the stem/progenitor cells are epithelial and/or mesenchymal stem/progenitor cells.5. The method of claim 1 , further comprising:(d) culturing the stem/progenitor cells under conditions allowing the cells to undergo clonal expansion.6. The method of claim 5 , further comprising:(e) culturing the stem/progenitor cells under conditions allowing the differentiation of said cells into ...

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Номер записи: 86
26-12-2013 дата публикации

UMBILICAL CORD PRODUCTS

Номер: US20130344163A1
Автор: Tan Ek Kia, TSENG Amy, Tseng Scheffer
Принадлежит:

Disclosed herein, in certain instances, are tissue grafts derived from UCAM. Further disclosed herein, in certain instances, are use for tissue grafts derived from UCAM. 123.-. (canceled)24. An umbilical cord graft derived from pre-frozen umbilical cord , wherein (a) water has not been removed from the umbilical cord graft , and (b) the umbilical cord graft is substantially free of a vein or artery.25. The umbilical cord graft of claim 24 , wherein the umbilical cord graft comprises umbilical cord amniotic membrane.26. The umbilical cord amniotic membrane graft of claim 25 , wherein the umbilical cord amniotic membrane comprises an epithelial layer claim 25 , a basement membrane claim 25 , a compact layer claim 25 , a fibroblast layer claim 25 , and a spongy layer.27. The umbilical cord graft of or claim 25 , substantially free of cells with metabolic activity.28. The umbilical cord graft of claim 24 , further comprising at least a portion of Wharton's Jelly.29. The umbilical cord graft of claim 24 , substantially free of at least a portion of Wharton's Jelly.30. The umbilical cord graft of claim 24 , wherein the umbilical cord graft is not lyophilized.31. The umbilical cord graft of claim 24 , wherein structural integrity of the umbilical cord graft is substantially preserved for at least 15 days after initial procurement.32. The umbilical cord graft of claim 24 , wherein natural biological activity of the umbilical cord graft is substantially preserved for at least 15 days after initial procurement.33. The umbilical cord graft of claim 24 , wherein the umbilical cord graft is cryopreserved claim 24 , sterilized claim 24 , or a combination thereof.34. The umbilical cord graft of claim 24 , wherein the umbilical cord graft is anti-inflammatory claim 24 , anti-scarring claim 24 , anti-angiogenic claim 24 , anti-adhesion claim 24 , or promotes wound healing.35. The umbilical cord graft of claim 24 , further comprising a supportive backing.36. The umbilical cord graft ...

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Номер записи: 87
26-12-2013 дата публикации

CULTURE MEDIA FOR DEVELOPMENTAL CELLS CONTAINING ELEVATED CONCENTRATIONS OF LIPOIC ACID

Номер: US20130344595A1
Автор: GARDNER David K., LARMAN Mark G., LINCK Donald
Принадлежит: Vitrolife Sweden AB

A composition and method for in vitro fertilization is provided which uses culture media comprising elevated concentrations of lipoic acid. More specifically, the invention provides culture media for developmental cells having a lipoic acid concentration of 5 μM to 40 μM. Culture media that include lipoic acid at concentrations within the identified range are able to provide blastocysts with increased survival, increased cell numbers, increased inner cell masses and/or increased percentage of the total mass made up by the inner cell compared to blastocysts cultured in a control medium. 1. A method for culturing developmental cells , the method comprising culturing a stem cell , gamete , zygote or embryo in a developmental cell culture medium , wherein the culture medium comprises:(a) water;(b) inorganic salts;(c) at least one energy source; and(d) lipoic acid, lipoic acid derivatives or a combination thereof at a concentration of about 5 μM to about 40 μM.2. The method of claim 1 , wherein the culture medium further comprises one or more amino acids.3. The method of claim 2 , wherein the concentration of inorganic salts in the culture medium is about 115 mM to about 140 mM claim 2 , the concentration of energy sources in the culture medium is about 5 mM to about 30 mM claim 2 , and the total concentration of amino acids in the culture medium is about 0.1 to 15 mM.4. The method of claim 1 , wherein the culture medium further comprises a chelating agent claim 1 , an antibiotic and hyaluronan.5. The method of claim 1 , wherein the culture medium supports the development of at least one of stem cells claim 1 , gametes claim 1 , zygotes or embryos from one developmental stage to another developmental stage.6. The method of claim 1 , wherein the stem cell claim 1 , gamete claim 1 , zygote or embryo is cultured under ambient oxygen conditions.7. The method of claim 1 , wherein the stem cell claim 1 , gamete claim 1 , zygote or embryo is cultured under reduced oxygen ...

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Номер записи: 88
26-12-2013 дата публикации

AUTOMATED SYSTEM FOR PRODUCING INDUCED PLURIPOTENT STEM CELLS OR DIFFERENTIATED CELLS

Номер: US20130345094A1
Автор: Chang Stephen, Eggan Kevin C., Noggle Scott, Solomon Susan
Принадлежит: New York Stem Cell Foundation

The invention provides an automated system for producing induced pluripotent stem cells (iPSCs) from adult somatic cells. Further, the system is used for producing differentiated adult cells from stem cells. The invention system is useful for isolating somatic cells from tissue samples, producing iPSC lines from adult differentiated cells by reprogramming such cells, identifying the pluripotent reprogrammed adult cells among other cells, and expanding and screening the identified reprogrammed cells. 1. An automated system for generating iPSCs , comprising:a cell plating unit for placing cells on a plate; andan induction unit for automated reprogramming of cells by contacting the cells on the plating unit with reprogramming factors to produce iPSCs.2. The system of claim 1 , further comprising:a sorting unit for selectively sorting and isolating the iPSCs produced by the induction unit by identifying iPS specific markers.3. The system of claim 2 , further comprising:an expansion unit for expanding the isolated iPSCs, and selecting the expanded iPSCs.4. The system of claim 1 , further comprising:a freezing unit for freezing the isolated iPSCs.5. The system of claim 1 , further comprising:a confluency check unit which checks the confluency of the iPSCs to determine whether the iPSCs have a confluency characteristic.6. The system of claim 5 , wherein the confluency check unit periodically checks the confluency of the iPSCs.7. The system of claim 5 , wherein the confluency check unit periodically checks the confluency of the iPSCs on a more frequent basis over time.8. The system of claim 5 , wherein the confluency check unit checks whether the confluency characteristic is within the range of 70% to 100%.9. The system of claim 1 , wherein the induction unit uses a viral vector to initiate reprogramming.10. The system of claim 9 , wherein the induction unit uses a retrovirus or a Sendai virus to initiate re-programming.11. The system of claim 1 , wherein the induction unit ...

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Номер записи: 89
09-01-2014 дата публикации

CULTURE VESSEL FOR FORMING EMBRYOID BODY

Номер: US20140011269A1
Автор: Sakura Takeshi, Tsukada Ryouhei
Принадлежит: SUMITOMO BAKELITE CO., LTD.

The present invention provides a culture vessel for forming an embryoid body that makes it possible to form an embryoid body of higher quality, more efficiently, at a higher level. The present invention relates to a culture vessel for forming an embryoid body that has two or more wells, in which the wells have a bottom having a cross section that looks like approximately a U shape when being observed in a vertical direction and an opening having an approximately circular shape, at least a curved portion of the inner surface of the bottom has low cell adhesive properties, a curvature radius (R′) of the inner surface of the bottom is from 1.0 mm to 3.5 mm, and the low cell adhesion surface is formed by being performed low-or-non cell adhesion treatment with a water-soluble resin. 1. A culture vessel for forming an embryoid body , comprising:a vessel structure having a plurality of wells,wherein each of the wells has a bottom having a cross section in an approximately U shape in a vertical direction and an opening having an approximately circular shape, each of the wells has a low cell adhesive property in at least a curved portion of an inner surface of the bottom, the inner surface of the bottom has a curvature radius of from 1.0 mm to 3.5 mm, and the inner surface is treated with a water-soluble resin such that the inner surface has the low cell adhesive property.3. The culture vessel for forming an embryoid body according to claim 1 , wherein each of the wells has at least one dent having a low cell adhesion surface inside a surface of a lowermost portion of the well.4. The culture vessel for forming an embryoid body according to claim 1 , wherein each of the wells has a low cell adhesive property on an entire inner surface of the well.5. The culture vessel for forming an embryoid body according to claim 2 , wherein each of the wells has at least one dent having a low cell adhesion surface inside a surface of a lowermost portion of the well.6. The culture vessel ...

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Номер записи: 90
23-01-2014 дата публикации

COMPOSITIONS AND METHODS FOR PROMOTING THE GENERATION OF DEFINITIVE ENDODERM

Номер: US20140024114A1
Автор: Borowiak Malgorzata, Chen Shuibing C., Fox Julia L., Maehr Rene, Melton Douglas A., Schreiber Stuart L., Tang Weiping
Принадлежит: President and Fellows of Harvard College

Certain embodiments disclosed herein are directed to a method of producing endoderm cells, such as definitive endoderm cells by exposing stem cells such as embryonic stem cells or induced pluripotent stem (iPS) cells to an effective amount of at least one compound described herein to differentiate the stem cells into the endoderm cells such as definitive endoderm cells. Differentiated endoderm cells produced by the methods disclosed herein can be differentiated into pancreatic epithelium, and other endoderm derivatives such as thymus, liver, stomach, intestine and lung. Another aspect of the present invention relates to a method of producing pancreatic progenitor cells, such as Pdx1-positive pancreatic progenitor cells by exposing endoderm cells, such as definitive endoderm cells to an effective amount of at least one compound described herein to differentiate the definitive endoderm cells into Pdx1-positive pancreatic progenitor cells. Kits and compositions comprising Pdx1-positive pancreatic progenitor produced using the methods are also described. 2. The method of claim 1 , wherein a Pdx1-positive pancreatic progenitor cell also expresses HNF6.4. The method of claim 3 , wherein the compound of Formula (II) is (2S claim 3 ,5S)-1 claim 3 ,2 claim 3 ,4 claim 3 ,5 claim 3 ,6 claim 3 ,8-Hexahydro-5-(hydroxymethyl)-1-methyl-2-(1-methylethyl)-3H-pyrrolo[4 claim 3 ,3 claim 3 ,2-gh]-1 claim 3 ,4-benzodiazonin-3-one ((−)-indolactam V).5. The method of claim 3 , further comprising isolating a population of Pdx1-positive pancreatic progenitor cells.6. The method of claim 5 , further comprising differentiating the population of Pdx1-positive pancreatic progenitor cells into a population of insulin producing cells.7. The method of claim 5 , further comprising differentiating the population of Pdx1-positive pancreatic progenitor cells into a population of cells having at least one characteristic of endogenous pancreatic β-cells.8. The method of claim 7 , wherein a cell with at ...

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Номер записи: 91
23-01-2014 дата публикации

Non-Static Suspension Culture of Cell Aggregates

Номер: US20140024116A1
Автор: Catherine M. Verfaillie, Kartik Subramanian, Wei-Shou Hu, Yonsil Park
Принадлежит: Katholieke Universiteit Leuven, University of Minnesota

The invention is directed to compositions of cell aggregates and methods for making and using the cell aggregates where the aggregates comprise cells that are not embryonic stem cells but can differentiate into cell types of at least two of ectodermal, endodermal, and mesodermal embryonic germ layers, e.g., stem cells.

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Номер записи: 92
23-01-2014 дата публикации

CELL REPROGRAMMING COMPOSITION COMPRISING REX1 AND AN INDUCED PLURIPOTENT STEM CELL PRODUCTION METHOD USING THE SAME

Номер: US20140024119A1
Автор: Cho Yee Sook, Son Mi Young
Принадлежит: Korea Research Institute of Bioscience and Biotechnology

A reprogramming inducing composition including the Rex1 protein or a nucleic acid molecule coding for the Rex1 protein for producing induced pluripotent stem cells from body cells or non-embryonic cells through a reprogramming process. A method for producing induced pluripotent stem cells by using the Rex1 is also disclosed. 1. A method for inducing reprogramming differentiated cells into induced pluripotent stem cells , comprising introducing a reprogramming-inducing factor comprising the Rex1 (reduced expression protein 1) protein or a nucleic acid molecule encoding the Rex1 protein into differentiated cells.2. The method according to claim 1 , wherein the nucleic acid molecule encoding the Rex1 protein is included in an expression vector.3. The method according to claim 1 , wherein the nucleic acid molecule encoding the Rex1 protein is messenger RNA (mRNA).4. The method according to claim 1 , wherein the reprogramming-inducing factor further comprises one or more proteins selected from the group consisting of Oct4 claim 1 , Sox2 claim 1 , KlF4 claim 1 , c-Myc claim 1 , Nanog claim 1 , and Lin-28 claim 1 , or one or more nucleic acid molecules encoding said proteins.5. The method according to claim 1 , wherein the cells are derived from human.6. The method according to claim 1 , wherein the differentiated cells are somatic cells or somatic stem cells.7. The method according to claim 6 , wherein the somatic stem cells are neural stem cells or mesenchymal stem cells.8. A method for producing induced pluripotent stem cells that are reprogrammed from differentiated cells claim 6 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) inducing reprogramming differentiated cells into induced pluripotent stem cells by the method according to , and'}(b) culturing the cells of step (a).9. The method according to claim 8 , further comprising (c) isolating embryonic stem cell-like colonies from the culture broth obtained in step (b).10. The method according to ...

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Номер записи: 93
30-01-2014 дата публикации

Methods and systems for harvesting cells

Номер: US20140030805A1
Автор: Barakf Zohar, Harel KASUTO, Nirit Drori-Carmi
Принадлежит: Pluristem Ltd

Methods for using vibration to harvest cells grown in 3 D culture are provided. The methods entail the application of force to cells attached to a 3 D matrix of sufficient amplitude, frequency, and duration to detach cells from the matrix and to flush the detached cells out of the matrix material. An apparatus for performing the methods of the invention as provided.

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Номер записи: 94
06-02-2014 дата публикации

Methods for reducing and/or preventing excessive cellular apoptosis

Номер: US20140037591A1
Автор: Banas Richard A., Wessel Howard C.
Принадлежит:

The invention is directed to novel methods for reducing the number of apoptotic cell deaths in a population of cells undergoing excessive cellular apoptosis. The invention is also directed to novel methods for preventing apoptotic cell death in a population of cells at risk for developing excessive cellular apoptosis. In particular, the invention is directed to novel methods for reducing or preventing excessive cellular apoptosis comprising exposing cells exhibiting or at risk for developing excessive cellular apoptosis to a novel cellular factor-containing composition called Amnion-derived Cellular Cytokine Solution (referred to herein as ACCS), which is obtained from the culturing of Amnion-derived Multipotent Progenitor (AMP) cells, or AMP cells. 1. A method for reducing the number of apoptosis-induced cell deaths in a population of cells undergoing excessive cellular apoptosis , the method comprising the step of contacting the population of cells undergoing excessive cellular apoptosis with a therapeutically effective dose of a composition selected from the group consisting of Amnion-derived Cellular Cytokine Solution (ACCS) and Amnion-derived Multipotent Progenitor (AMP) cells , such that the number of apoptosis-induced cell deaths is reduced.2. The method of wherein the ACCS comprises physiologic concentrations of VEGF claim 1 , TGFβ2 claim 1 , Angiogenin claim 1 , PDGF claim 1 , TIMP-1 and TIMP-2 claim 1 , wherein the physiologic concentration is about 5.0-16 ng/mL for VEGF claim 1 , about 3.5-4.5 ng/mL for Angiogenin claim 1 , about 100-165 pg/mL for PDGF claim 1 , about 2.5-2.7 ng/mL for TGFβ2 claim 1 , about 0.68 μg/mL for TIMP-1 claim 1 , and about 1.04 μg/mL for TIMP-2.3. The method of wherein the excessive cellular apoptosis occurs in a subject as a result of a condition selected from the group consisting of an ischemic condition claim 1 , a radiation-induced injury claim 1 , and an injury to nervous tissue.4. The method of wherein the excessive ...

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Номер записи: 95
13-02-2014 дата публикации

METHODS FOR CULTURING STEM AND PROGENITOR CELLS

Номер: US20140044696A1
Автор: Bamdad Cynthia
Принадлежит: MINERVA BIOTECHNOLOGIES CORPORATION

The present application describes a method of culturing, expanding or growing stem or stem-like cells or induced pluripotent stem cells on a surface, including attaching the cells to the surface through a ligand that binds to the surface and the cells. 123-. (canceled)241. A method of harvesting cells from cells grown according to the method of claim , comprising adding a competing molecule that binds to the ligand so that the cells are released from binding to the ligand or the surface.251. A method of harvesting cells from cells grown according to the method of claim , comprising cleaving a linker bound to the surface that is directly or indirectly attached to the cells , so that the cells are released from the surface.26. A method of identifying state of differentiation of cells comprising using anti-MUC1* antibody to bind to the cells , wherein positive signal for anti-MUC1* antibody indicates puripotent cell state , and cells showing binding to non-clipped MUC1 indicates differentiated cell state.27. The method according to claim 26 , comprising separating cells from a mixed population of stem and stem-like cells or induced pluripotent stem cells and newly differentiating cells claim 26 , comprising using anti-MUC1* antibody to bind to the cells claim 26 , wherein positive signal for anti-MUC1* antibody indicates a pluripotent cell state claim 26 , and cells showing binding to non-clipped MUC1 indicates differentiated cell state.28. The method according to claim 27 , further comprising contacting the cells with antibodies to stem or stem-like cell or induced pluripotent stem cell markers claim 27 , wherein positive signal for a stem or stem-like cell or induced pluripotent stem cell marker indicates the presence of pluripotent stem cell state.29. The method according to claim 28 , wherein the cells are contacted with anti-MUC1* and anti-Tra 1-81 claim 28 , anti-Tra 1-60 claim 28 , SSEA3 or SSEA4 antibodies.30. A method of detecting cancer stem cells using anti- ...

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Номер записи: 96
13-02-2014 дата публикации

METHODS AND COMPOSITIONS FOR PROMOTING SURVIVAL AND PROLIFERATION OF ENDOTHELIAL CELLS AND STIMULATING ANGIOGENESIS

Номер: US20140045260A1
Автор: Rafii Shahin, SEANDEL Marco, Zhang Fan
Принадлежит: Cornell Research Foundation, Inc.

The present invention relates to adenovirus E4ORF1 gene and to endothelial cells engineered to express the E4ORF1 gene. The present invention also relates to uses of the E4ORF1 gene, and cells expressing the E4ORF1 gene, and to compositions comprising the E4ORF1 gene, or comprising cells expressing the E4ORF1 gene. 185.-. (canceled)86. A method of culturing cancer cells , stem cells or progenitor cells , the method comprising:(a) obtaining or generating a population of E4ORF1-expressing endothelial cells; and(b) culturing the E4ORF1-expressing endothelial cells in the same culture vessel with cancer cells, stem cells or progenitor cells.87. The method of claim 86 , wherein the E4ORF1-expressing endothelial cells form a feeder cell layer in a culture vessel claim 86 , and wherein the cancer cells claim 86 , stem cells or progenitor cells are placed on the feeder cell layer.88. The method of claim 86 , wherein the endothelial cells are primary endothelial cells.89. The method of claim 86 , wherein the endothelial cells are human umbilical vein endothelial cells (HUVECs).90. The method of claim 86 , wherein the stem cells are hematopoietic stem cells or embryonic stem cells.91. The method of claim 86 , wherein the stem cells are hematopoietic stem cells and wherein the endothelial cells and the hematopoietic stem cells are grown in the absence of serum.92. The method of claim 86 , wherein the stem cells are embryonic stem cells and wherein the endothelial cells and the embryonic stem cells are grown in the absence of serum.93. A population of cancer cells claim 86 , stem cells or progenitor cells obtained using the method of .94. A method of culturing cancer cells claim 86 , stem cells or progenitor cells claim 86 , the method comprising:(a) obtaining or generating a population of E4ORF1-expressing endothelial cells;(b) culturing the E4ORF1-expressing endothelial cells in a culture vessel;(c) collecting conditioned medium from the culture vessel; and(d) adding the ...

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Номер записи: 97
13-02-2014 дата публикации

POSTPARTUM CELLS DERIVED FROM UMBILICAL CORD TISSUE, AND METHODS OF MAKING AND USING THE SAME

Номер: US20140045263A1
Автор: Gosiewska Anna, HARMON ALEXANDER M., Harris Ian R., Kihm Anthony J., Messina Darin J., Mistry Sanjay, Seyda Agnieszka, Yi Chin-Feng
Принадлежит: DEPUY SYNTHES PRODUCTS, LLC.

Cells derived from human umbilical cords are disclosed along with methods for their therapeutic use. Isolation techniques, culture methods and detailed characterization of the cells with respect to their cell surface markers, gene expression, and their secretion of trophic factors are described. 1. A method of enhancing the yield of cells isolated from human umbilical cord tissue , the method comprising the steps of:(a) obtaining umbilical cord tissue;(b) removing substantially all of the blood from the tissue to yield umbilical tissue substantially free of blood;(c) dissociating the umbilical tissue substantially free of blood by mechanical dissociation such as mincing;(d) digesting the dissociated tissue with a mixture of enzymes comprising a metalloprotease, neutral protease and mucolytic enzyme;(e) isolating umbilicus-derived cells from the digested tissue;(f) resuspending the isolated umbilicus-derived cells in a growth medium; and(g) culturing the isolated umbilicus-derived cells for about 10 to 100 hours to obtain a substantially homogenous population of isolated umbilicus-derived cells, wherein the isolated umbilicus-derived cells are capable of self-renewal and expansion in culture and have the potential to differentiate into cells of other phenotypes.2. The method of claim 1 , wherein the removing step comprises removal of free or clotted blood by one or more of washing claim 1 , suctioning claim 1 , blotting claim 1 , centrifugal separation claim 1 , or enzymatic removal.3. The method of claim 1 , wherein the metalloprotease enzyme is collagenase.4. The method of claim 1 , wherein the neutral protease enzyme is dispase.5. The method of claim 1 , wherein the mucolytic enzyme is hyaluronidase.6. The method of claim 1 , wherein the digestion step comprises incubating the dissociated tissue with the mixture of enzymes at about 37° C.7. The method of claim 1 , wherein the digestion step comprises incubating the dissociated tissue with the mixture of enzymes ...

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Номер записи: 98
13-02-2014 дата публикации

Methods of culturing cells in a medium comprising transforming growth factor beta 1 and basic fibroblast growth factor

Номер: US20140045266A1
Автор: Joseph Itskovitz-Eldor, Michal Amit
Принадлежит: Technion Research and Development Foundation Ltd

The present invention is of methods of establishing and propagating human embryonic stem cell lines using feeder cells-free, xeno-free culture systems and stem cells which are capable of being maintained in an undifferentiated, pluripotent and proliferative state in culture which is free of xeno contaminants and feeder cells.

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Номер записи: 99
20-02-2014 дата публикации

STEM CELL CULTURE METHOD

Номер: US20140050704A1
Автор: Forsyth Nicholas, Kumar Deepak, Yang Ying
Принадлежит: KEELE UNIVERSITY

Methods for culturing pluripotent stem cells on fiber scaffolds are provided which result in the expansion of the number of stem cells without loss of pluripotency. Cells obtained by such methods, implants containing such cells and medical methods using such cells are also disclosed. 1. A method of culturing human embryonic stem cells (hESC) whilst maintaining stemness , the method comprising seeding hESC onto an uncoated fiber scaffold and incubating the hESC loaded fiber scaffold in a gas atmosphere in which the percentage of oxygen is less than the percentage of oxygen in air.2. The method of wherein the fiber scaffold comprises nanofibers.3. The method of wherein the fiber scaffold comprises electrospun fibers.4. The method of wherein the percentage of oxygen is in the range 0.25% to 20%.5. The method of wherein the percentage of oxygen is in the range 0.5% to 10%.6. The method of wherein the percentage of oxygen is about 2%.7. The method of wherein the fibers have a diameter of between 50 nm and 3000 nm.8. The method of wherein the fibers have a diameter of between 100 nm and 300 nm.9. The method of wherein the fiber scaffold comprises aligned fibers.10. The method of wherein the fibers comprise a synthetic polymer.11. The method of wherein the fibers comprise poly-ε-caprolactone (PCL).12. The method of wherein the pluripotent stem cells are incubated for 10 days without loss of pluripotency.13. The method of further comprising the step of inducing the pluripotent stem cells to differentiate.14. The method of wherein the inducing the pluripotent stem cells to differentiate comprises adding differentiation media to the fibers seeded with pluripotent stem cells.15. The method of wherein inducing the pluripotent stem cells to differentiate comprises removing the pluripotent stem cells from the fibers before adding differentiation medium.16. A method of preparing a population of human embryonic stem cells (hESC) that are suitable for clinical use comprising ...

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Номер записи: 100