СРЕДА ДЛЯ КУЛЬТИВИРОВАНИЯ КЛЕТОК БЕЗ БЕЛКОВ И БЕЗ СЫВОРОТКИ

Изобретение относится к биотехнологии и может быть использовано при культивировании клеток млекопитающих, а также для получения белка из клеток млекопитающих. Питательная среда представляет собой синтетическую среду с добавлением гидролизата сои в количестве от 0,1 до 100 г/л, причем по меньшей мере 40% гидролизата имеет м.м. ≤500 Да. Среда может также содержать буфер, стабилизатор окисления и т.д., а синтетической средой является среда DMEM/HAM F12, 199 или RPMI. При выращивании клеток в данной среде увеличивается продукция как рекомбинантных клеток, так и их продуктивность (увеличение выхода белка). Кроме того, среда является универсальной в плане выбора метода культивирования клеток млекопитающего. 3 н. и 11 з.п. ф-лы, 5 ил., 7 табл.

КОРРЕКТИРОВКА PH ДЛЯ УЛУЧШЕНИЯ РАЗМОРАЖИВАНИЯ БАНКОВ КЛЕТОК

Изобретение относится к области биотехнологии, а именно к улучшению размораживания банков клеток, замораживанию клеток яичника китайского хомячка (СНО), композиции для замораживания клеток СНО и банку клеток. Способ включает замораживание клеток CHO в среде для замораживания, которая содержит буферный раствор и DMSO, при рН среды для замораживания, доведенном до от 7,5 до 8,5 перед замораживанием. Изобретение позволяет повысить жизнеспособность и скорость роста клеток при оттаивании. 6 н. и 63 з.п. ф-лы, 6 ил., 1 пр.

ПОПУЛЯЦИЯ СТВОЛОВЫХ КЛЕТОК ШЕЙКИ МАТКИ ЧЕЛОВЕКА И ЕЕ ПРИМЕНЕНИЕ

Изобретение относится к области клеточной биологии и биотехнологии, в частности к способу выделения мезенхимальных стволовых клеток шейки матки, не являющихся опухолевыми, которые экспрессируют клеточные маркеры CD29, CD44, CD73, CD105 и CD90 и не экспрессируют клеточные маркеры CD117, CD133, HLA-DR, TRA-81, CD45, CD34 и CD31, применению выделенной ткани шейки матки и способу получения кондиционированной среды. Способ включает приготовление клеточной суспензии из ткани шейки матки, полученной десквамацией шейки матки, выделение клеток из указанной клеточной суспензии, инкубацию указанных клеток в подходящей среде для культивирования клеток и в условиях, которые обеспечивают пролиферацию клеток и отбор мезенхимальных стволовых клеток, где указанные мезенхимальные стволовые клетки шейки матки экспрессируют клеточные маркеры CD29, CD44, CD73, CD105 и CD90 и не экспрессируют клеточные маркеры CD117, CD133, HLA-DR, TRA1-81, CD45, CD34 и CD31. Далее осуществляют инкубацию данных клеток в среде ...

НЕ СОДЕРЖАЩАЯ ЖИВОТНЫХ БЕЛКОВ СРЕДА ДЛЯ КУЛЬТИВИРОВАНИЯ КЛЕТОК

Изобретение относится к области биотехнологии, конкретно к получению питательных сред для культивирования клеток, и может быть использовано для значительного снижения вариаций продукции рекомбинантных белков, которые имеют место при культивировании клеток с использованием разных партий коммерчески доступного соевого гидролизата. Получают не содержащую животных белков среду для культивирования клеток путем дополнения сред, не содержащих животные белки, гидролизатом сои и дополнительно - биогенным амином в диапазоне 1-18 мг/л. Полученную среду используют для получения рекомбинантных белков, посредством процессов культивирования соответствующих клеток. Изобретение позволяет понизить контагиозность продукта, а также увеличить эффективность роста и продуктивность клеток. 3 н. и 9 з.п. ф-лы, 9 ил.

МЕТОДИКА КУЛЬТИВИРОВАНИЯ КЛЕТОК

Изобретение относится к области биотехнологии. Изобретение представляет собой среду для культивирования клеток, содержащую тирозин в концентрации от 4 до 50 мМ и поливиниловый спирт (PVA) в концентрации от 0,5 до 10 г/л и способ культивирования клеток, включающий приведение клеток млекопитающего в контакт со средой для культивирования клеток эмбриональной почки человека (293), клеток почки новорожденного хомячка (BHK), клеток яичника китайского хомячка (CHO), мышиных клеток Сертоли, клеток почки африканской зеленой мартышки (VERO-76), клеток рака шейки матки человека (HeLa), клеток почки собаки, клеток печени крысы линии Buffalo, клеток легкого человека, клеток печени человека, клеток опухоли молочной железы мыши, клеток TRI, клеток MRC 5, клеток FS4 или линии гепатомы человека (Hep G2). Таким образом, разработана улучшенная среда для культивирования клеток с повышенной растворимостью тирозина для культивирования клеток млекопитающих в высокой плотности и для оптимального получения белков ...

ШТАММ КЛЕТОК CHO-SE-9/4 - ПРОДУЦЕНТ ХИМЕРНОГО АНТИТЕЛА ПРОТИВ ЭРИТРОПОЭТИНА ЧЕЛОВЕКА И ХИМЕРНОЕ АНТИТЕЛО, ПРОДУЦИРУЕМОЕ ДАННЫМ ШТАММОМ

Настоящее изобретение относится к области иммунологии. Предложено антитело, способное специфически связываться с эритропоэтином человека. Также рассмотрен штамм-продуцент такого антитела. Данное изобретение может найти дальнейшее применение в выделении и/или очистке рекомбинантного эритропоэтина человека методом иммуноаффинной хроматографии. 2 н.п. ф-лы, 6 ил., 6 пр.

АМНИОТИЧЕСКИЕ АДГЕЗИВНЫЕ КЛЕТКИ

СПОСОБ ПОЛУЧЕНИЯ СЛИТОГО БЕЛКА, ИМЕЮЩЕГО FC-ДОМЕН IgG

ПОПУЛЯЦИЯ СТВОЛОВЫХ КЛЕТОК ШЕЙКИ МАТКИ ЧЕЛОВЕКА И ЕЕ ПРИМЕНЕНИЕ

Obtaining cellular and DNA material from human female reproductive system for various purposes including ovarian and endometrial cancer detection

Efficient harvesting of cells and DNA material from female reproductive system is made possible by processing gelatinous part of cervical mucus. Rinsing may be used to separate the gelatinous part of cervical mucus from the bulk of cervical cells. The harvested cells and DNA material may be used for diagnostic purposes including ovarian and endometrial cancer detection.

Human ovarian mesothelial cells and methods of isolation and uses thereof

Reduction of high molecular weight species, acidic charge species, and fragments in a monoclonal antibody composition

Alterations in bioreactor cell culture feeding to an extended or continuous feed following an initial period of no feeding reduces the level of high molecular weight, acid charge, and fragment species of monoclonal antibodies expressed in the culture, and enhances the level of afucosylated species of monoclonal antibodies expressed in the culture. Regular fucose infusions following an initial period of no feed media infusion reduces the level of afucosylated species of monoclonal antibodies expressed in the culture. Cell culture manipulation may be used to modulate the level of species of monoclonal antibodies.

Modulating lactogenic activity in mammalian cells

The present disclosure relates to methods, cells and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interests, where the cells (e.g, Chinese Hamster Ovary (CHO) cells) have modulated lactogenic activity. The present disclosure also relates to methods and compositions for modulating pyruvate kinase muscle (PKM) expression (e.g, PKM-l expression) in a mammalian cell to thereby reduce or eliminate the lactogenic activity of the cell, as well compositions comprising a cell having reduced or eliminated lactogenic activity and methods of using the same.

DELIVERY OF PAYLOADS TO STEM CELLS

The present disclosure relates to a method of targeting stems cells, in particular non-apoptotic stem cells, employing a GLA domain, capable of binding surface exposed phosphatidyl serine.

SEED TRAIN PROCESSES AND USES THEREOF

Provided herein are seed train processes and methods of producing a recombinant protein that include the use of these seed train processes.

REGULATING ORNITHINE METABOLISM TO MANIPULATE THE HIGH MANNOSE GLYCOFORM CONTENT OF RECOMBINANT PROTEINS

The present invention relates to a method for manipulating the high mannose glycoform content of recombinant glycoproteins by regulating ornithine metabolism during cell culture.

COMPOSITIONS AND METHODS FOR ENHANCING BIOENERGETIC STATUS IN FEMALE GERM CELLS

Compositions and methods comprising bioenergetic agents for restoring the quality of aged oocytes, enhancing oogonial stem cells or improving derivatives thereof (e.g., cytoplasm or isolated mitochondria) for use in fertility- enhancing procedures, are described.

METHODS FOR ENHANCED PRODUCTION OF BONE MORPHOGENETIC PROTEINS

Methods and processes for improved recombinant protein production are provided. The methods are useful for production of growth factors, particularly those of the TGF-.beta. superfamily, including bone morphogenetic proteins (BMPs), such as BMP-2. Suitable host cells are cultured in media where iron is present at a concentration of at least 2.25 µM and if pyridoxal is present, it makes up less than 55 % of the molar concentration of vitamin B6 in the media.

CHROMOSOMAL LANDING PADS AND RELATED USES

Provided herein are methods for stable integration and/or expression of one or more recombinant polynucleotides in a host cell. The recombinant polynucleotides are typically integrated into the host genome at some native chromosomal integration sites. The integration can be mediated by homologous recombination or by using a hybrid recombinase targeting the specific chromosomal locations. The native chromosomal integration sites in the host cells, which support stable integration and strong transcription activities of foreign genes, are present within or adjacent to specific genes in the CHO genome, ankyrin 2 gene (Ank2), cleavage and polyadenylation specific factor 4 gene (Cpsf4), C-Mos gene, and Nephrocystin-l/Mal gene. Also provided are methods and nucleic acid molecules for inserting site-specific recombination sequences (chromosomal landing pads) into these specific chromosomal locations. Further provided are engineered host cells which contain chromosomal landing pads or transgenes ...

ADDITION OF IRON TO IMPROVE CELL CULTURE

The present invention provides, among other things methods of increasing cell density, viability and/or titer in a cell culture including steps of adding a composition comprising iron to the cell culture.

METHODS AND COMPOSITIONS FOR THE CLINICAL DERIVATION OF AN ALLOGENIC CELL AND THERAPEUTIC USES

Various cells, stem cells, and stem cell components, including associated methods of generating and using such cells are provided. In one aspect, for example, an isolated cell that is capable of self-renewal and culture expansion and is obtained from a subepithelial layer of a mammalian umbilical cord tissue. Such an isolated cell expresses at least three cell markers selected from CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and does not express at least three cell markers selected from CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.

METHODS FOR PREPARING PROTEIN CULTIVATION OF MAMMALIAN CELLS

COMPOSITIONS AND METHODS INCREASE CONDENSATE BIOENERGETIC STATE OF FEMALE GERM CELLS

METABOLICALLY OPTIMISED CELL CULTURE

MODULATION OF LIPID METABOLISM FOR PRODUCING PROTEINS

PRODUCTION OF ANTIBODIES FOR CONJUGATION FROM CULTURES OF CELLS CHO

METHOD OF PREPARATION Of a FARMING SYSTEM Of ENDOMETRE AUTOLOGUE FOR CO-CULTURE ENDOMETRE-EMBRYON

célula isolada, população isolada de células, matriz ou estrutura sintética permanente ou decelularizada degradável, e , método para tratar um indivíduo

PROCEDURE OF CULTURE OF CELLS OF MAMMAL FOR THE PRODUCTION OF PROTEINS

Procedimientos y procesos para la produccion de proteínas, en particular de glucoproteínas, mediante el cultivo de células animales o de mamífero, preferentemente, pero no limitados a, cultivos celulares semicontinuos. En un aspecto, los procedimientos comprenden la adicion de compuesto glucocorticoide durante el periodo de cultivo. La adicion de compuesto glucocorticoide mantiene una viabilidad alta de los cultivos celulares y puede proporcionar un aumento en la titulacion final de producto proteico y una calidad alta de producto proteico, tal como se determina, por ejemplo, por medio del contenido de acido siálico en la proteína producida. La consistencia en la calidad del producto proteico, producido de manera recombinante, es un requisito para la aprobacion regulatoria del mismo para ser empleado como agente terapéutico y/o profiláctico.

METHODS AND COMPOSITIONS FOR THE CLINICAL DERIVATION OF AN ALLOGENIC CELL AND THERAPEUTIC USES

METHODS FOR ENHANCED PRODUCTION OF BONE MORPHOGENETIC PROTEINS

METHOD FOR CULTURING ADHESIVE CELLS, CULTURE VESSEL, AND METHOD FOR PRODUCING PROTEIN

The present invention is a method for culturing adherent cells that can allow adherent cells to survive and grow even in a suspended state, a culture vessel that is formed using an alicyclic structure-containing polymer, and includes a liquid medium, and adherent cells that survive in the liquid medium in a suspended state, and a method for producing a protein. The adhesive cells can be grown in a method for culturing adherent cells comprising bringing adherent cells into contact with an alicyclic structure-containing polymer formed article to grow the adherent cells in a state in which the adherent cells are suspended in a liquid medium. 1. A method for culturing adherent cells comprising bringing adherent cells into contact with an alicyclic structure-containing polymer formed article to grow the adherent cells in a state in which the adherent cells are suspended in a liquid medium.2. The method for culturing adherent cells according to claim 1 , wherein the adherent cells are genetically engineered cells that express a foreign gene.3. The method for culturing adherent cells according to claim 1 , wherein the adherent cells are CHO cells.4. The method for culturing adherent cells according to claim 1 , wherein the adherent cells are cells that can express a foreign gene.5. The method for culturing adherent cells according to claim 1 , wherein the adherent cells that are suspended in the liquid medium form a cell mass.6. A culture vessel that is formed using an alicyclic structure-containing polymer claim 1 , the culture vessel comprising a liquid medium claim 1 , and adherent cells that survive in the liquid medium in a suspended state.7. The culture vessel according to claim 6 , wherein the adherent cells form a cell mass.8. A method for producing a protein comprising bringing recombinant cells that can express a foreign gene that encodes a physiologically active protein into contact with an alicyclic structure-containing polymer formed article when the ...

Primary cultures of normal and tumoral human ovarian epithelium

The present invention relates to primary cultures or established cell lines of ovarian epithelium origin which are substantially equivalent to the cells from the original clinical material from which they are derived and can serve as a powerful model in numerous types of studies including the elaboration of a patient-based tumor profile, thereby permitting a more precise and personalized design of an efficacious therapeutic regimen for cancer therapy. The invention further relates to a method to derive primary cell cultures from benign and malignant ovarian tissue and from ascites.

CELL CULTURE METHODS FOR EXPRESSING ADAMTS PROTEIN

The present invention provides culture mediums that are useful for the expression of ADAMTS proteins, such as ADAMTS13. Methods for the expression and purification of ADAMTS proteins are also provided. In some embodiments, the mediums and methods of the invention are useful for the expression of ADAMTS proteins having high specific activities. Also provided are ADAMTS, e.g., ADAMTS13, protein compositions with high specific activities, which are expressed and purified according to the methods provided herein.

MGAT1-Deficient Cells for Production of Vaccines and Biopharmaceutical Products

Mannosyl (alpha-1,3)-glycoprotein beta-1,2-N-Acetylglucosaminyltransferase (Mgat1)-deficient cell lines and methods for use of same for producing human immunodeficiency virus (HIV) envelope glycoprotein polypeptides or fragment thereof with terminal mannose-5 glycans are provided.

Method for Obtaining Female Germline Stem Cells from Follicular Aspirates

Provided is a method for obtaining female germline stem cells from follicular aspirates.

Method for slowing down aging in women by prolonging ovarian function

The main characteristic of method is that before the onset of menopause in women at age 30-45 years, part of the cortical and medulla layers of one or both ovaries is surgically resected and cryopreserved (frozen), and in perimenopause beginning, these tissues are autotransplanted subcutaneously in a form of suspension or small ovarian pieces prepared from thawed tissues fragments of their own ovaries. This “return” is repeatedly periodically each 4-6 months for 10-15 years. That helps to obtain normal level of ovarian antiage factors in blood till age 60-65 years, thus slowing aging.

Tetracycline-inducible expression systems

The invention provides inducible promoter systems and their components incorporating components of a tetracycline operon. By coordinating expression of different transcriptional units in these systems as a result of selection of promoters and/or linking the units into the same DNA molecule, these systems can achieve higher levels of expression of coding segments of interest, increased differential levels of expression between on- and off-states, and/or greater responsiveness to inducing agents than conventional systems.

Human embryo co-culture system and uses thereof

Methods of in vitro oocyte development

Methods of preparing ovarian tissue for primordial follicle growth are presented comprising the steps: providing an ovarian tissue sample comprising cortical tissue and stromal tissue; removing damaged tissue from the ovarian tissue sample where present; removing excess stromal tissue from the ovarian tissue sample where present; and then mechanically stretching the ovarian tissue sample along at least one dimension of the ovarian tissue sample, such that the size of the ovarian tissue sample along the at least one dimension is increased by at least 10%. Methods of growing viable oocyte in vitro, and methods of preparing individual ovarian follicles for growth are also presented.

Methods for modulating production profiles of recombinant proteins

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also 5 disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant.

Seed train processes and uses thereof

Abstract Provided herein are seed train processes and methods of producing a recombinant protein that include the use of these seed train processes.

EGFR x CD28 multispecific antibodies

The present invention provides multispecific antibodies that bind to EGFR and CD28 (EGFRxCD28) as well as anti-EGFR antibodies. Such antibodies may be combined with a further therapeutic agent such as an anti-PD1 antibody. Methods for treating cancers ( ...

METHODS AND SYSTEMS FOR ENDOMETRIAL TREATMENT

STIMULATION OF OVARIAN FOLLICLE DEVELOPMENT AND OOCYTE MATURATION

Methods are provided for stimulating ovarian follicles in a mammal through activation of the mTor signaling pathway.

METHOD FOR PRODUCING FUSION PROTEIN HAVING IGG FC DOMAIN

The present invention relates to a method for preparing a fusion protein having an IgG Fc domain and, specifically, to a method for preparing a fusion protein having an IgG Fc domain, the method additionally comprising a step of culturing cells, which produce the fusion protein, at a decreased culture temperature, thereby increasing cell growth and cell viability so as to increase fusion protein productivity and inhibiting aggregate generation so as to improve quality and production yield.

METHODS FOR MODULATING PRODUCTION PROFILES OF RECOMBINANT PROTEINS

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also 5 disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant.

REDUCTION OF HIGH MOLECULAR WEIGHT SPECIES, ACIDIC CHARGE SPECIES, AND FRAGMENTS IN A MONOCLONAL ANTIBODY COMPOSITION

Alterations in bioreactor cell culture feeding to an extended or continuous feed following an initial period of no feeding reduces the level of high molecular weight, acid charge, and fragment species of monoclonal antibodies expressed in the culture, and enhances the level of afucosylated species of monoclonal antibodies expressed in the culture. Regular fucose infusions following an initial period of no feed media infusion reduces the level of afucosylated species of monoclonal antibodies expressed in the culture. Cell culture manipulation may be used to modulate the level of species of monoclonal antibodies.

MICROFLUIDIC METHODS AND CARTRIDGES FOR CELL SEPARATION

The invention discloses a method for selecting cells depending on their level of displaying and preferably secreting a protein of interest from a population of heterogeneously expressing cells, comprising: (a) contacting said cells with magnetic beads coated with an affinity group to the said cells, (b) mixing the said magnetic beads with the cells to capture the cells displaying/secreting the protein of interest, (c) performing at least one washing step to remove the non-captured cells, and (d) recovering the cells to which that magnetic beads have bound.

AMNION DERIVED ADHERENT CELLS

Provided herein are novel angiogenic cells from amnion, referred to as amnion derived adherent cells, and populations of, and compositions comprising, such cells. Further provided herein are methods of obtaining such cells and methods of using the cells in the treatment of individuals.

AMNION DERIVED ADHERENT CELLS

Provided herein are novel angiogenic cells from amnion, referred to as amnion derived adherent cells, and populations of, and compositions comprising, such cells. Further provided herein are methods of obtaining such cells and methods of using the cells in the treatment of individuals.

COMPOSITIONS AND METHODS FOR ENHANCING BIOENERGETIC STATUS IN FEMALE GERM CELLS

REGULATION OF METABOLISM ORNITHINE FOR CONTROL OF CONTENT OF HIGH MANNOZNYKhGLIKOFORM RECOMBINANT PROTEINS

METHODS FOR PREPARING PROTEIN CULTIVATION OF MAMMALIAN CELLS

METABOLICALLY OPTIMISED CELL CULTURE

CHO DG44 (Chinese Hamster Ovary) culture medium and application thereof

Separation and extraction method of laying duck small follicular theca cells

USE OF N-ACETYL-5-METHOXYTRYPTAMINE OR ANALOGUES THEREOF, FOR PROMOTING THE MECHANISM OF IMPLANTATION OF THE EMBRYO AND RELATED COMPOSITIONS AND CULTURE MEDIA

발효 시스템

... 본 발명은 현재 기존의 발효 전략보다 이점을 지닌 신규한 세포 배양 및 세포 및/또는 세포-유래된 생성물 생산 공정에 관한 것이다. 본 발명에 따른 공정 및 방법은 현재 적용되는 공정(배치, 유가 또는 관류 전략)에 비해 이점을 지닌 대안적인 생산 공정으로서, 일시적인 생산 플랫폼에 대한 매우 생육가능하고 대사적으로 활성인 진핵생물 세포의 효율적인 공급을 위해 그리고 일시적인 발현 시스템을 위한 후속 사용 또는 바이러스 또는 슈도바이러스에 의해 또는 세포 비함유 시스템에서 감염을 위한 대사적으로 매우 활성인 바이오매스를 생성하기 위해 이용될 수 있다.

CELL CULTURE MEDIA AND METHODS OF ANTIBODY PRODUCTION

Hücrelerin proteinsiz veya serumsuz yerleştirilmesi için ortam

INTERPENETRATING BIOMATERIAL MATRICES AND USES THEREOF

The present invention relates to matrices (e.g., fibrin-alginate matrices; fibrin-alginate-matrigel matrices) for culture of cells, organs (e.g., ovary or fragment thereof), cells and cell aggregates (e.g., ovarian follicles, embryoid bodies), and tissues. In some embodiments, protease inhibitors e.g., aprotinin) are used to prevent the degradation of fibrin.

Chromosomal Landing Pads and Related Uses

Provided herein are methods for stable integration and/or expression of one or more recombinant polynucleotides in a host cell. The recombinant polynucleotides are typically integrated into the host genome at some native chromosomal integration sites. The integration can be mediated by homologous recombination or by using a hybrid recombinase targeting the specific chromosomal locations. The native chromosomal integration sites in the host cells, which support stable integration and strong transcription activities of foreign genes, are present within or adjacent to specific genes in the CHO genome, ankyrin 2 gene (Ank2), cleavage and polyadenylation specific factor 4 gene (Cpsf4), C-Mos gene, and Nephrocystin-1/Mal gene. Also provided are methods and nucleic acid molecules for inserting site-specific recombination sequences (chromosomal landing pads) into these specific chromosomal locations. Further provided are engineered host cells which contain chromosomal landing pads or transgenes ...

ENDOMETRIAL REGENERATIVE CELLS FOR TREATMENT OF TRAUMATIC BRAIN INJURY

The use of endometrial regenerative cells (ERC) and other endometrial originating cells for the treatment of traumatic brain injury is disclosed. In one embodiment a patient is administered a population of CD90 positive, CD105 positive, allogeneic regenerative cells subsequent to a brain injury. Cell concentration, frequency of administration, and route of administration may be determined based on extent of injury, inflammatory response and endogenous stem cell mobilization. In one embodiment, a patient suffering from traumatic brain injury is administered a dose of 100 million Endometrial Regenerative Cells intravenously at a rate of 1 million cells per minute in a volume of 100 ml of saline. 1. A method of treating a neurological injury comprising administering into a patient an effective amount of endometrial regenerative cells.2. The method of claim 1 , wherein said neurological injury is acute or chronic.3. The method of claim 2 , wherein said acute neurological injury is selected from the group consisting of: a) traumatic brain injury; b) stroke; and c) intracerebral hemorrhage.4. The method of claim 1 , wherein said endometrial regenerative cells originate from the endometrium and said cell expresses a marker selected from the group consisting of: CD29 claim 1 , CD41a claim 1 , CD44 claim 1 , CD90 claim 1 , and CD105 claim 1 , and have an ability to proliferate at a rate of 0.5-1.5 doublings per 24 hours in a growth medium.5. The method of claim 1 , wherein said cell further expresses a marker selected from the group consisting of: NeuN claim 1 , CD9 claim 1 , CD62 claim 1 , CD59 claim 1 , Actin claim 1 , GFAP claim 1 , NSE claim 1 , Nestin claim 1 , CD73 claim 1 , SSEA-4 claim 1 , hTERT claim 1 , Oct-4 claim 1 , and tubulin.6. The method of claim 1 , wherein said cell further expresses a marker selected from hTERT and Oct-4 claim 1 , but does not express a STRO-1 marker claim 1 , and has an ability to undergo cell division in less than 24 hours in a growth ...

Human ovarian mesothelial cells and methods of isolation and uses thereof

The invention discloses a substantially pure population of human ovarian mesothelial cells and methods of isolating and culturing the ovarian mesothelial cells. By carefully manipulating the microenvironment of the ovarian mesothelial cells, multiple passages are attainable wherein the ovarian mesothelial cells are capable of becoming ovary surface epithelial cells or granulosa cells. In addition, several methods of use of human ovarian mesothelial cells are disclosed herein.

CULTURE MEDIA COMPRISING N-ACYL-X-GLUTAMINE DIPEPTIDES

Disclosed herein is a cell culture media containing L-glutamine from a set of N-acylated dipeptides Acyl-X-Q, and L-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource), wherein the variables X, Q, Acyl, R, n(Acyl-X-Q) and n(Qsource) are defined in the general disclosure. Processes of using the cell culture media are also described herein. 1. A cell culture media , comprisingL-glutamine from a set of N-acylated dipeptides Acyl-X-Q; andL-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource),wherein:X is defined as an L-amino acid;Q is defined as L-glutamine attached via an amide bond to L-amino acid X;Acyl is defined as a C1-C7-acyl moiety attached via an amide bond to the amino-terminus of L-amino acid X;R is defined to be in the range of 0,03 to 20;n(Acyl-X-Q) is the total amount of substance of L-glutamine contained in the set of N-acylated dipeptides Acyl-X-Q in the culture media;n(Qsource) is the total amount of substance of L-glutamine contained in the set of other glutamine sources Qsource in the culture media; andthe constituents of the set of other L-glutamine sources (Qsource) are selected from the following: Free L-glutamine, dipeptides Y-Q, or mixtures thereof, wherein Y is defined as one of the 20 genetically encoded L-amino acids, wherein Q is defined as L-glutamine attached via an amide bond to L-amino acid Y, and the amide bond connecting Y and Q in dipeptide Y-Q is a regular backbone amide bond involving the carboxy terminus of amino acid Y and the amino terminus of glutamine Q.2. The cell culture media according to claim 1 , wherein the cell culture media are serum free.3. The cell culture media according to claim 1 , wherein the cell culture media are chemically defined.4. The cell culture media according to claim 1 , wherein the only constituent of the set of other L-glutamine sources Qsource is dipeptide Y-Q claim 1 , wherein Y is Alanine.5. ...

IMPROVED MEDIA FOR THE EXPRESSION OF RECOMBINANT VITAMIN K-DEPENDENT PROTEINS

The present invention relates to a method for increasing the activity and/or the yield of a recombinant vitamin K-dependent protein expressed in cell culture. The present invention further relates to uses and compositions of matter.

CHINESE HAMSTER OVARY CELL LINES

We provide a Chinese Hamster Ovary (CHO) cell which is capable of higher protein sialylation compared to a wild type Chinese Hamster Ovary cell, such as in the presence of functional GnT 1, in which the CHO cell is obtainable by selection with Ricinus communis agglutinin I (RCA-I).

PROCESS OF PRODUCTION WITH CONTROLLED COPPER IONS

The present invention provides a method of cell culture comprising adding a cell-containing seed medium to an initial medium and starting to culture the cell in the initial medium, wherein the initial medium has an organism-derived culture medium additive added thereto and the amount of the C-terminal amidated species in the produced protein is controlled by the copper content of the initial medium at the start of cell culture.

METHODS OF IMPROVING PROTEIN TITER IN CELL CULTURE

Methods of improving recombinant protein titer and cell titer in cell culture using cell culture media having reduced impurities are provided, and well as cell culture media having reduced impurities that can used for the production of a recombinant protein and cells with improved titer. The cell culture media having reduced impurities comprises a HEPES buffer, and the reduced impurities are HEPES related impurities. In certain aspects, methods and media improve protein titer, cell growth, and/or viable cell density.

Marker genes for oocyte competence

Cumulus cell (CC) gene expression is being explored as an additional method to morphological scoring to choose the embryo with the highest chance to pregnancy. The present invention relates to a novel method of identifying biomarker genes for evaluating the competence of a mammalian oocyte in giving rise to a viable pregnancy after fertilization, based on the use of live birth and embryonic development as endpoint criteria for the oocytes to be used in an exon level analysis of potential biomarker genes. The invention further provides CC-expressed biomarker genes thus identified, as well as prognostic models based on the biomarker genes identified using the methods of the present invention.

CELL STRAIN FOR PRODUCING BIOSIMILAR DRUG OF USTEKINUMAB AND PRODUCTION METHOD THEREFOR

Provided are a cell strain for producing a biosimilar drug of Ustekinumab and a production method therefor. Specifically, provided is a Chinese hamster ovary cell S cell strain. The cell strain expresses a full human monoclonal antibody directed against the P40 subunit shared by human IL-12 and human IL-23. The fully human monoclonal antibody directed against the P40 subunit shared by human IL-12 and human IL-23 is a biosimilar drug of Ustekinumab, which not only exhibits high consistency with Ustekinumab in pre-clinical research, but also passes pharmacokinetic bioequivalence and safety similarity evaluation in clinical research. The biosimilar drug of Ustekinumab is the first one that has entered clinical trials in China, is the only one that has completed the I stage clinical trial, and is also one of the biosimilar drugs of Ustekinumab, which has the fastest progress in new drug application in the world.

METHODS FOR DETERMINING DEVELOPMENTAL STAGE OF HUMAN CUMULUS CELLS

PROCUREMENT, ISOLATION AND CRYOPRESERVATION OF ENDOMETRIAL/MENSTRUAL CELLS

СПОСОБ ПОЛУЧЕНИЯ БЕЛКА

Изобретение относится к области молекулярной биологии и генетической инженерии. Предложен способ получения представляющего интерес белка, включающему введение вектора экспрессии белка, который включает генный фрагмент, содержащий ДНК, кодирующую представляющий интерес белок, и ген селектируемого маркера, а также транспозонные Tol1 или Tol2 последовательности на обоих концах генного фрагмента, в суспензионную клетку млекопитающего СНО, адаптированную к суспензионному культивированию, или клетку PER.C6, клетки крысиной миеломы YB2/3HL.Р2.G11.16Ag.20 (или также называемой YB2/0), или клетку мышиной миеломы NS0, адаптированную к суспензионному культивированию; интегрирование генного фрагмента, вставленного между парой транспозонных последовательностей, в хромосому клетки млекопитающего для получения клетки млекопитающего, способной экспрессировать представляющий интерес белок; и суспензионное культивирование клетки млекопитающего; при этом суспензионная клетка млекопитающего способна экспрессировать ...

УЛУЧШЕННЫЕ ЭУКАРИОТИЧЕСКИЕ КЛЕТКИ ДЛЯ ПОЛУЧЕНИЯ БЕЛКА И СПОСОБЫ ИХ ПОЛУЧЕНИЯ

Изобретение относится к биотехнологии, в частности описаны клетки млекопитающих и линии клеток млекопитающих с уменьшенной нагрузкой остатков возбудителей ранее перенесенных вирусных/ретровирусных инфекций и способы получения и применения таких клеток. 4 н. и 32 з.п. ф-лы, 22 ил., 3 табл., 1 пр.

СПОСОБ АДРЕСНОГО ВОЗДЕЙСТВИЯ НА ЭКЗОСОМЫ

ПРИМЕНЕНИЕ N-АЦЕТИЛ-5-МЕТОКСИТРИПТАМИНА ИЛИ ЕГО АНАЛОГОВ ДЛЯ АКТИВАЦИИ МЕХАНИЗМА ИМПЛАНТАЦИИ ЭМБРИОНА, А ТАКЖЕ СООТВЕТСТВУЮЩИЕ КОМПОЗИЦИИ И КУЛЬТУРАЛЬНЫЕ СРЕДЫ

... 1. N-ацетил-5-метокситриптамин или его аналог для применения в области медицины или ветеринарии при вспомогательной репродукции для профилактики неудачного исхода имплантации в матку путем местного применения эффективного количества у самки млекопитающего, нуждающейся в таком лечении.2. N-ацетил-5-метокситриптамин или его аналог по п. 1, где указанный аналог выбран из агомелатина, 6-гидроксимелатонина, серотонина, 5-гидрокситриптофана или их производных.3. N-ацетил-5-метокситриптамин или его аналог по п. 1, где указанной самкой млекопитающего является женщина, страдающая бесплодием или множественными выкидышами.4. N-ацетил-5-метокситриптамин или его аналог по п. 1, где указанное местное применение осуществляют посредством орошения эндометрия, промывания матки или промывания эндометрия.5. N-ацетил-5-метокситриптамин или его аналог по п. 1, где указанное местное применение проводят путем единственного применения во время извлечения яйцеклеток.6. N-ацетил-5-метокситриптамин или его аналог ...

КОМПОЗИЦИЯ НА ОСНОВЕ ГИДРОЛИЗАТА БЕЛКА ГОРОХА ДЛЯ ПОВЫШЕНИЯ СПЕЦИФИЧЕСКОЙ ПРОДУКТИВНОСТИ КЛЕТОЧНЫХ ЛИНИЙ ПРОДУЦЕНТОВ И СПОСОБ ЕЁ ПОЛУЧЕНИЯ

Группа изобретений относится к биотехнологии. Композиция для повышения специфической продуктивности клеточных линий-продуцентов антител на основе СНО клеток содержит следующие исходные компоненты на 1 л: L-глутамин в количестве 1,168 г, Pluronic F-68 в количестве 1 г, инсулин в количестве 0,01 г, трансферрин в количестве 0,0055 г, селенит натрия в количестве 0,0000067 г, гидролизат белков гороха в количестве 0,06 г и среду IMDM - остальное до 1 л. Способ получения композиции включает следующие этапы. Готовят белки путем смешивания исходных компонентов в виде их растворов в буфере с исходной концентрацией 0,7 мг/мл. Экстрагируют белки и центрифугируют. Осуществляют качественный и количественный анализ и подвергают гидролизу с папаином. Останавливают реакцию с последующим количественным анализом. Выпавший осадок удаляют, а раствор экстрагированного белка фильтруют на мембране для отсечения компонентов гидролизата с молекулярной массой выше 5 кДа. Группа изобретений обеспечивает повышение ...

КОМПОЗИЦИЯ НА ОСНОВЕ ГИДРОЛИЗАТА БЕЛКА ПОДСОЛНЕЧНИКА ДЛЯ ПОВЫШЕНИЯ СПЕЦИФИЧЕСКОЙ ПРОДУКТИВНОСТИ КЛЕТОЧНЫХ ЛИНИЙ ПРОДУЦЕНТОВ И СПОСОБ ЕЁ ПОЛУЧЕНИЯ

Группа изобретений относится к биотехнологии. Композиция для повышения специфической продуктивности клеточных линий-продуцентов антител на основе СНО клеток содержит следующие исходные компоненты на 1 л: L-глутамин в количестве 1,168 г, Pluronic F-68 в количестве 1 г, инсулин в количестве 0,01 г, трансферрин в количестве 0,0055 г, селенит натрия в количестве 0,0000067 г, гидролизат белка подсолнечника в количестве 0,07 г и среду IMDM - остальное до 1 л. Способ получения композиции включает следующие этапы. Готовят белки путем смешивания исходных компонентов в виде их растворов в буфере с исходной концентрацией 0,7 мг/мл. Экстрагируют белки и центрифугируют. Осуществляют качественный и количественный анализ и подвергают гидролизу с папаином. Останавливают реакцию с последующим количественным анализом. Выпавший осадок удаляют, а раствор экстрагированного белка фильтруют на мембране для отсечения компонентов гидролизата с молекулярной массой выше 5 кДа. Группа изобретений обеспечивает повышение ...

ПРОЦЕССЫ КУЛЬТИВИРОВАНИЯ В СИСТЕМЕ ПОСЕВНЫХ ФЕРМЕНТЕРОВ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

MEDIUM FOR PROTEIN AND SERUM-FREE THE CELL CULTURE

HUMAN EPITHELIAL CELLS OF MÜLLERIAN DUCTUS DESCENT, PROCEDURE FOR YOUR ISOLATION AND USES OF IT

HUMAN EIERSTOCKMESOTHELIALZELLEN AND PROCEDURES FOR THEIR ISOLATION AND USE

Animal protein-free media for cultivation of cells

Amnion derived adherent cells

Procurement, isolation and cryopreservation of endometrial/menstrual cells

Cell culture medium for ADAMTS protein expression

The present invention provides culture mediums that are useful for the expression of ADAMTS proteins, such as ADAMTS13. Methods for the expression and purification of ADAMTS proteins are also provided. In some embodiments, the mediums and methods of the invention are useful for the expression of ADAMTS proteins having high specific activities. Also provided are ADAMTS, e.g., ADAMTS13, protein compositions with high specific activities, which are expressed and purified according to the methods provided herein.

PD-1 antibodies

The present invention provides antibodies that bind human programmed cell death 1 (PD-1), and may be useful for treating cancer alone and in combination with chemotherapy and other cancer therapeutics.

Human uterine cervical stem cell population and uses thereof

The present invention relates to a method for isolating stem cells comprising preparing a cell suspension from uterine cervix tissue, to the stem cells isolated by said method, and to the conditioned medium obtained from the culture of said stem cells. The invention also encompasses the use of said stem cells or conditioned medium for treating or preventing cancer, precancerous lesions, inflammatory diseases, autoimmune diseases, chronic pathologies or infectious diseases, diseases associated to tissue loss, or for use in diagnostic, prognostic or treatment of fertility disorders, as well as for cosmetic treatment.

Microcarrier perfusion culturing methods and uses thereof

Provided herein are methods of culturing a mammalian cell and various methods that utilize these culturing methods.

Method for producing fusion protein having IgG Fc domain

The present invention relates to a method for preparing a fusion protein having an IgG Fc domain and, specifically, to a method for preparing a fusion protein having an IgG Fc domain, the method additionally comprising a step of culturing cells, which produce the fusion protein, at a decreased culture temperature, thereby increasing cell growth and cell viability so as to increase fusion protein productivity and inhibiting aggregate generation so as to improve quality and production yield.

Methods of selecting and designing safer and more effective anti-CTLA-4 antibodies for cancer therapy

The present invention relates to compositions of anti-CTLA-4 antibodies that bind to the human CTLA4 molecule and their use in cancer immunotherapy and for the reduction of autoimmune side effects compared to other immunotherapeutic agents.

Integration sites in CHO cells

The present invention relates to the identification of a genomic integration site for heterologous polynucleotides in Chinese Hamster Ovary (CHO) cells resulting in high RNA and/or protein production. More specifically it relates to CHO cells comprising at least one heterologous polynucleotide stably integrated into the S100A gene cluster of the CHO genome and to methods for the production of said CHO cells. Further, the invention relates to a method for the production of a protein of interest using said CHO cell and to the use of said CHO cell for producing a protein of interest at high yield. Integration within these specific target regions leads to reliable, stable and high yielding production of an RNA and/or protein of interest, encoded by the heterologous polynucleotide.

Cell culture process

A cell culture medium comprising tyrosine at a concentration of at least 3 mM and polyvinylalcohol (PVA).

Methods for preparing antibodies with a defined glycosylation pattern

The present disclosure relates to a process for preparing antibodies with a defined glycosylation pattern, in particular antibodies with a glycan terminating in an N-acetylglucosamine. The antibodies of the disclosure are suitable for use in a process to conjugate a payload thereto. The disclosure also extends to molecules obtained and obtainable from the process disclosed herein, novel molecules and intermediates, compositions comprising said molecules and uses of the molecules and compositions, particularly in treatment, for example in the treatment of cancer.

ENDOMETRIAL STEM CELLS AND METHODS OF MAKING AND USING SAME

The invention provides pluripotent stem cells and methods for making and using pluripotent stem cells. Pluripotent stem cells, among other things, can differentiate into various cell lineages in vitro, ex vivo and in vivo. Pluripotent stem cells, among other things, can also be used to produce conditioned medium. 1. A human pluripotent stem cell that expresses a marker selected from CD29 , CD41a , CD44 , CD90 , and CD105 , and having an ability to proliferate at a rate of 0.5-1.5 doublings per 24 hours in a growth medium.2. The human pluripotent stem cell of claim 1 , wherein said cell further expresses a marker selected from NeuN claim 1 , CD9 claim 1 , CD62 claim 1 , CD59 claim 1 , Actin claim 1 , GFAP claim 1 , NSE claim 1 , Nestin claim 1 , CD73 claim 1 , SSEA-4 claim 1 , hTERT claim 1 , Oct-4 claim 1 , and tubulin.3. The human pluripotent stem cell of claim 1 , wherein said cell further expresses a marker selected from hTERT and Oct-4 claim 1 , but does not express a STRO-1 marker claim 1 , and has an ability to undergo cell division in less than 24 hours in a growth medium.4. The human pluripotent stem cell of claim 1 , wherein said cell further expresses a STRO-1 marker claim 1 , and has an ability to proliferate at a rate of 0.5-0.9 doublings per 24 hours in a growth medium.5. (canceled)6. The human pluripotent stem cell of claim 1 , wherein said cell produces matrix metalloprotease 3 (MMP3) claim 1 , matrix metalloprotease 10 (MMP10) claim 1 , GM-CSF claim 1 , PDGF-BB or angiogenic factor ANG-2.7. The human pluripotent stem cell of claim 1 , wherein said cell is derived or originates from endometrium claim 1 , endometrial stroma claim 1 , endometrial membrane claim 1 , or menstrual blood.818.-. (canceled)19. The human pluripotent stem cell of claim 1 , wherein said cell is capable of differentiating into an adipogenic claim 1 , endothelial claim 1 , hepatic claim 1 , osteogenic claim 1 , neural claim 1 , pancreatic or myocytic cell lineage.20. The human ...

METHODS AND COMPOSITIONS FOR THE CLINICAL DERIVATION OF AN ALLOGENIC CELL AND THERAPEUTIC USES

Various cells, stem cells, and stem cell components, including associated methods of generating and using such cells are provided. In one aspect, for example, an isolated cell that is capable of self-renewal and culture expansion and is obtained from a subepithelial layer of a mammalian umbilical cord tissue. Such an isolated cell expresses at least three cell markers selected from CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and does not express at least three cell markers selected from CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR. 1. An isolated cell obtained from a subepithelial layer of a mammalian umbilical cord tissue capable of self-renewal and culture expansion;wherein the isolated cell expresses at least three cell markers selected from the group consisting of CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105; andwherein the isolated cell does not express at least three cell markers selected from the group consisting of CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.2. The isolated cell of claim 1 , wherein the isolated cell expresses CD29 claim 1 , CD73 claim 1 , CD90 claim 1 , CD166 claim 1 , SSEA4 claim 1 , CD9 claim 1 , CD44 claim 1 , CD146 claim 1 , and CD105.3. The isolated cell of claim 1 , wherein the isolated cell does not express CD45 claim 1 , CD34 claim 1 , CD14 claim 1 , CD79 claim 1 , CD106 claim 1 , CD86 claim 1 , CD80 claim 1 , CD19 claim 1 , CD117 claim 1 , Stro-1 claim 1 , and HLA-DR.4. The isolated cell of claim 1 , wherein the isolated cell is positive for SOX2.5. The isolated cell of claim 1 , wherein the isolated cell is positive for OCT4.6. The isolated cell of claim 1 , wherein the isolated cell is positive for SOX2 and OCT4.7. The isolated cell of claim 1 , wherein the wherein the isolated cell is capable of differentiation into a cell type selected from the group consisting of adipocytes claim 1 , chondrocytes claim 1 , osteocytes claim 1 , ...

Methods Of Gene Amplification And Expression

Disclosed are methods relating to amplification and expression of a nucleic acid sequence encoding a polypeptide of interest in recombinant cells, and cell lines and polypeptides produced from such methods. The methods disclosed herein permit the amplification of cell lines that express a polypeptide of interest in a relatively short period of time through the use of a bioreactor. 1. A method for producing a recombinant cell line that expresses a polypeptide of interest , comprising (a) culturing a host cell in a bioreactor in media , wherein the host cell is transfected with (i) a nucleic acid sequence encoding a polypeptide of interest and (ii) a nucleic acid sequence encoding an amplifiable marker; and (b) adding an increasing amount of an amplification agent to the media to produce the recombinant cell line.2. The method of wherein the nucleic acid sequence encoding a polypeptide of interest and the nucleic acid sequence encoding an amplifiable marker are contained on a single vector.3. The method of wherein the amplifiable marker encodes a DHFR protein.4. The method of wherein the host cells are mammalian cells.5. The method of wherein the host cells are Cl-JO cells.6. The method of wherein the polypeptide of interest is an antibody.7. The method of wherein the bioreactor is a perfusion bioreactor.8. The method of wherein the increasing amount of the amplification agent is added to the media when % cell viability is increasing or stable.9. A method for producing a polypeptide of interest claim 1 , which comprises culturing host cells transfected with (i) a nucleic acid sequence encoding a polypeptide of interest and (ii) a nucleic acid sequence encoding an amplifiable marker in a bioreactor in media containing increasing amounts of an amplification agent claim 1 , thereby expressing the polypeptide of interest.10. The method of wherein the nucleic acid sequence encoding a polypeptide of interest and the nucleic acid sequence encoding an amplifiable marker are ...

METHOD FOR PREPARING AQUEOUS SOLUTION CONTAINING CULTURE MEDIUM AND CHELATING AGENT

Provided are a method for preparing a highly versatile aqueous solution having remarkably improved membrane filterability, which can be stably membrane-filtered in a short time, an aqueous solution prepared by the preparation method, a method for culturing cells using the aqueous solution which is prepared by the preparation method, a method for producing a physiologically active substance using the culturing method, a physiologically active substance produced by the method for producing a physiologically active substance, a method for performing membrane filtration of the aqueous solution which is prepared by the preparation method of the aqueous solution, a method for improving membrane filterability of the aqueous solution, and a method for producing the physiologically active substance by preparing the aqueous solution, performing membrane filtration of the aqueous solution, and then culturing cells using the resulting aqueous solution. The present invention relates to a method for preparing an aqueous solution, characterized by addition of a chelating agent. 1. A method for preparing an aqueous solution comprising a culture medium and a chelating agent , wherein the chelating agent is added to the aqueous solution prior to the final pH adjustment of the aqueous solution.2. The method for preparing an aqueous solution according to claim 1 , wherein the chelating agent is one or more selected from citric acid claim 1 , malic acid claim 1 , ethylenediaminetetraacetic acid claim 1 , ethylenediaminetetraacetic acid iron(III) sodium salt claim 1 , sialic acid claim 1 , and salts or hydrates thereof.3. The method for preparing an aqueous solution according to or claim 1 , wherein the culture medium is a powdered culture medium claim 1 , a liquid culture medium claim 1 , or a slurry culture medium.4. The method for preparing an aqueous solution according to claim 3 , wherein the powdered culture medium further includes one or more selected from metal salts claim 3 , ...

SERUM-FREE MAMMALIAN CELL CULTURE MEDIUM, AND USES THEREOF

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× ...

Human uterine cervical stem cell population and uses thereof

The present invention relates to a method for isolating stem cells comprising preparing a cell suspension from uterine cervix tissue, to the stem cells isolated by said method, and to the conditioned medium obtained from the culture of said stem cells. The invention also encompasses the use of said stem cells or conditioned medium for treating or preventing cancer, precancerous lesions, inflammatory diseases, autoimmune diseases, chronic pathologies or infectious diseases, diseases associated to tissue loss, or for use in diagnostic, prognostic or treatment of fertility disorders, as well as for cosmetic treatment.

METHOD FOR DYNAMICALLY REMOVING RECOMBINANT HUMAN NERVE GROWTH FACTOR PRECURSOR BY HYDROPHOBIC INTERACTION CHROMATOGRAPHY

A method for removing precursors in recombinant human nerve growth factor (rhNGF) by hydrophobic interaction chromatography (HIC) is provided, where a Chinese hamster ovary (CHO) cell culture is processed by column chromatography for preliminary purification, and the pretreated sample obtained therefrom is further processed in a HIC column by washing the sample and then eluting the HIC column. 1. A method for removing a precursor in recombinant human nerve growth factor (rhNGF) by hydrophobic interaction chromatography (HIC) , comprising:1) processing a Chinese hamster ovary (CHO) cell culture by column chromatography for preliminary purification, thereby obtaining a product for further chromatographic separation;2) loading the product obtained from step 1) in a HIC column, washing the product with a washing buffer and discarding the eluate containing the precursor; and3) eluting the HIC column used in step 2) using an elution buffer, thereby obtaining a purified rhNGF product.2. The method of claim 1 , wherein said washing in step 2) comprises using a washing buffer comprising an alcohol and NaCl.3. The method of claim 2 , wherein said washing buffer satisfies each of the following conditions:A. having higher electrical conductivity than the chromatography elution buffer in step 3);B. having a lower alcohol content than the chromatography elution buffer in step 3); andC. being within a same pH range as the first product obtained from step 1).4. The method of claim 2 , wherein said alcohol is ethanol.5. The method of claim 2 , wherein said washing buffer has an ethanol content of from 4% to 6% by weight.6. The method of claim 2 , wherein said washing buffer has an NaCl content of from 200 to 400 mM.7. The method of claim 1 , wherein said washing in step 2) comprises using a washing volume which is determined by the following linear equation of a peak area of the first product in the column chromatography in step 1):{'br': None, 'washing volume (in the unit of CV)=8. ...

In Vivo Methods for Enhancing Bioenergetic Status in Female Germ Cells

In vivo methods using bioenergetic agents for restoring the quality of aged oocytes, enhancing oogonial stem cells or improving derivatives thereof (e.g., cytoplasm or isolated mitochondria) for use in fertility-enhancing procedures, are described. 1. A method of improving fertility in a female mammalian subject in need thereof , said method comprising administering to the female mammalian subject one or more bioenergetics agents in an amount effective to enhance the bioenergetic status of an oocyte or an oogonial stem cell (OSC) of the female mammalian subject , thereby improving fertility in the female mammalian subject.2. The method of claim 1 , wherein the bioenergetic agent is selected from the group consisting of a CD38 inhibitor claim 1 , a nicotinamide adenine dinucleotide (NAD) precursor and combinations thereof.3. The method of claim 2 , wherein the CD38 inhibitor is a compound selected from the group consisting of 1-[(2-Acetoxyethoxy)methyl]-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1-[(2-Benzyloxyethoxy)methyl]-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1{[2-(4-Methoxy-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1-{[2-(4-Phenoxy-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1-{[2-(4-Nitro-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1-{[2-(3-Trifluoromethyl-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1-{[2-(8′-Quinolyloxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride claim 2 , 1 claim 2 ,2-Dimethoxy-ethylene-bis-N claim 2 ,N′-3-(aminocarbonyl)-pyridinium dichloride claim 2 , 1 claim 2 ,4-Dimethoxy-butylene-bis-N claim 2 ,N′-3-(aminocarbonyl)-pyridinium dichloride claim 2 , 1 claim 2 ,4-Dimethoxy-butyne-bis-N claim 2 ,N′-3-(aminocarbonyl)-pyridinium dichloride claim 2 , 1 claim 2 ,4-Dimethoxy-hexamethylene-bis-N claim 2 ,N′-3-(aminocarbonyl)-pyridinium dichloride claim 2 , (E)-1-{[4-(8′-Quinolyloxy)but-2-enyloxy]methyl}-3-( ...

CHEMICALLY DEFINED CELL CULTURE MEDIA ADDITIVE

The present invention relates to an optimal medium for growing a cell line auxotrophic for tetrahydrofolate (THF) and producing a desired material in the cell with high efficiency, in particular, the present invention provides a method for enhancing cell growth by adding tetrahydrofolate (THF), or a precursor or derivative thereof into a chemical composition cell medium. 1. A cell culture medium comprising a chemically defined medium and tetrahydrofolate (THF) or a precursor or derivative thereof.2. The cell culture medium according to claim 1 , wherein the cells are auxotrophic for tetrahydrofolate.3. The cell culture medium according to claim 1 , wherein the cells auxotrophic for tetrahydrofolate have a functionally deleted or impaired DHFR gene.4. The cell culture medium according to claim 2 , wherein the cells auxotrophic for tetrahydrofolate are selected from the group consisting of mammalian cells claim 2 , insect cells claim 2 , plant cells claim 2 , and fungal cells.5. The cell culture medium according to claim 4 , wherein the mammalian cells are CHO cells.6. The cell culture medium according to claim 5 , wherein the CHO cells are selected from the group consisting of UKB25 claim 5 , DUK22 claim 5 , DUK51 claim 5 , DUK-D1 claim 5 , S1 claim 5 , DUK51-R1 claim 5 , DUK51-R2 claim 5 , DUK22-R1 claim 5 , DUK22-R2 claim 5 , DXBA claim 5 , DXE11 claim 5 , DXC11 claim 5 , DXB11 claim 5 , and DUKX.7. The cell culture medium according to claim 5 , wherein the CHO cells are selected from the group consisting of UA2 claim 5 , UA4 claim 5 , UA21 claim 5 , UA41 claim 5 , DU5 claim 5 , DU11 claim 5 , DG21 claim 5 , DG22 claim 5 , DG23 claim 5 , DG24 claim 5 , DG41 claim 5 , DG42 claim 5 , DG43 claim 5 , DG44 claim 5 , DG45 claim 5 , and DG46.8. The cell culture medium according to claim 1 , wherein the THF or precursor or derivative thereof is present at a concentration in the range of 0.1 to 160 mg/L.9. The cell culture medium according to claim 1 , wherein the THF ...

MODULATING LACTOGENIC ACTIVITY IN MAMMALIAN CELLS

The present disclosure relates to methods, cells and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interests, where the cells (e.g., Chinese Hamster Ovary (CHO) cells) have modulated lactogenic activity. The present disclosure also relates to methods and compositions for modulating pyruvate kinase muscle (PKM) expression (e.g., PKM-1 expression) in a mammalian cell to thereby reduce or eliminate the lactogenic activity of the cell, as well compositions comprising a cell having reduced or eliminated lactogenic activity and methods of using the same. 1. A mammalian cell having reduced or eliminated lactogenic activity , wherein the expression of a pyruvate kinase muscle (PKM) polypeptide isoform is knocked down or knocked out , and wherein the PKM polypeptide isoform comprises a PKM-1 polypeptide isoform.2. The mammalian cell of claim 1 , wherein the expression of a PKM-2 polypeptide isoform is knocked down or knocked out.3. The mammalian cell of claim 1 , wherein the cell is a CHO cell.4. The mammalian cell of claim 1 , comprising a nucleic acid sequence encoding a product of interest.5. The mammalian cell of claim 4 , wherein the product of interest comprises: (i) a protein; (ii) a recombinant protein; (iii) an antibody or an antigen-binding fragment thereof; or (iv) a combination thereof.6. The mammalian cell of claim 5 , wherein (i) the antibody is a multispecific antibody or an antigen-binding fragment thereof or (ii) the antibody consists of a single heavy chain sequence and a single light chain sequence or antigen-binding fragments thereof.7. The mammalian cell of claim 4 , wherein the nucleic acid sequence is integrated in the cellular genome of the mammalian cell at a targeted location.8. The mammalian cell of claim 1 , wherein the lactogenic activity of the mammalian cell is less than about 50% or less than about 20% of the ...

METHODS AND SYSTEMS FOR ENDOMETRIAL TREATMENT

Methods of treating endometrial dysfunction are described including retrieval of endometrial cells including endometrial stem cells from a fertile donor and transplantation of endometrial cells or tissue into a host suffering from endometrial dysfunction. In certain embodiments, methods include genetic modification, tissue culturing, or cryopreservation of the endometrial cells prior to transplantation. In various aspects, endometrial cells are retrieved from an individual during peak fertility and banked for autologous transplantation at a later time. 1. A method for enhancing endometrial function and fertility , the method comprising:obtaining an endometrial stem cell from a donor; andtransplanting the endometrial stem cell into a host uterus.2. The method of wherein the donor is also the host.3. The method of further comprising cryopreserving the endometrial stem cell.4. The method of wherein the endometrial stem cell is obtained within a 5 year period after the donor has given birth.5. The method of wherein the endometrial stem cell is obtained by endometrial biopsy.6. The method of wherein the endometrial stem cell is obtained by isolation from menstrual fluid.7. The method of wherein the endometrial stem cell is transplanted during the proliferative phase of the uterine cycle of the host.9. The method of claim 1 , wherein the transplantation step further comprises:injection of the endometrial stem cell into endometrium of the host uterus.10. The method of claim 1 , further comprising genetically modifying the endometrial stem cell before transplantation.11. The method of claim 10 , wherein the genetic modification comprises using a clustered regularly interspaced short palindromic repeat (CRISPR) nuclease.12. The method of claim 10 , wherein the genetic modification comprises augmenting expression of a protein linked to endometrial receptivity.13. The method of claim 1 , further comprising screening the donor for endometrial function.14. The method of claim 1 ...

MICROCARRIER PERFUSION CULTURING METHODS AND USES THEREOF

Provided herein are methods of culturing a mammalian cell and various methods that utilize these culturing methods.

METHOD, DEVICE AND KIT FOR MASS CULTIVATION OF CELLS USING POLYIMIDE POROUS MEMBRANE

The present invention pertains to a method for the mass cultivation of cells, and a cell cultivation device and kit. The present invention further pertains to a continuous cell cultivation method and a continuous cell cultivation device in which a carrier is used. 1. A mass cell culturing method including:(1) applying cells to a porous polyimide film, and(2) applying the porous polyimide film to which the cells have been applied, to a cell culture medium and performing culturing.2. The method according to claim 1 , wherein two or more porous polyimide films layered either above and below or left and right are used in the cell culture medium.3. The method according to or claim 1 , wherein the porous polyimide films are:i) folded,ii) wound into a roll,iii) connected as sheets or fragments by a filamentous structure, oriv) bound into a rope,to be suspended or fixed in the cell culture medium in the cell culturing vessel.4. The method according to any one of to claim 1 , wherein in the culturing of step (2) claim 1 , all or some of the porous polyimide films are not in contact with the liquid phase of the cell culture medium.5. The method according to any one of to claim 1 , wherein in the culturing of step (2) claim 1 , the total volume of the cell culture medium in the cell culturing vessel is 10 claim 1 ,000 times or less of the total sum of the porous polyimide film volume including the cell survival zone.6. The method according to any one of to claim 1 , wherein in the culturing of step (2) claim 1 , the total volume of the cell culture medium in the cell culturing vessel is 100 times or less of the total sum of the porous polyimide film volume including the cell survival zone.7. The method according to any one of to claim 1 , wherein the culturing in step (2) is carried out in a system in which a cell culture medium is continuously or intermittently supplied to a cell culturing vessel from cell culture medium supply means installed outside of the cell culturing ...

BOTTOM SECTION FOR BEING CONNECTED TO AN ASSEMBLY WITH PLATE SETTLER, AND ASSEMBLY WITH PLATE SETTLER

This disclosure relates to a bottom section for being connected to an assembly for separating a solid component from a fluid. The assembly includes an inclined plate settler with at least one sedimentation channel for letting a solid component to be separated settle, said plate settler comprising a lower portion and an upper portion, wherein said at least one sedimentation channel extends from the lower portion to the upper portion. The bottom section is configured to be connected to the lower portion of the inclined plate settler. The bottom section comprises at least one inlet channel for feeding a fluid comprising the solid component to be separated to the plate settler, and at least one collection channel for collecting a settled solid component descending from the at least one sedimentation channel. Said at least one inlet channel and said at least one collection channel are fluidly separated from each other, said inlet channel and said collection channel being connectable to said at least one sedimentation channel, to form fluid connections between said at least one inlet channel and said at least one sedimentation channel and between said at least one collection channel and said at least one sedimentation channel, respectively. 1. A bottom section for being connected to an assembly for separating a solid component from a fluid , said assembly including an inclined plate settler with at least one sedimentation channel for letting a solid component to be separated settle , said plate settler comprising a lower portion and an upper portion , wherein said at least one sedimentation channel extends from the lower portion to the upper portion , wherein the bottom section is configured to be connected to the lower portion of the inclined plate settler ,the bottom section comprising at least one inlet channel for feeding a fluid comprising the solid component to be separated to the plate settler, and at least one collection channel for collecting a settled component ...

A METHOD OF PREDICTING RELATIVE FED BATCH PRODUCTION TITER OF A PANEL OF CLONALLY-DERIVED PRODUCER CELLS

“A rapid, high throughput, method of predicting relative fed batch performance of a panel of clonal cells derived from a single host cell line”. A computer implemented method of predicting relative fed batch production titer of a panel of clonal producer cells derived from a single parental host cell population comprises the steps of assaying a level of growth of each clone in the presence and absence of at least three individual chemical cell stressors, comparing the level of growth of each clone in the presence and absence of the at least three chemical cell stressors to provide a normalised production titer response for each clone in each stressed microenvironment, and inputting a clone-specific production titer response profile comprising the normalised production titer response for each clone in each stressed microenvironments into a computational model. The computational model is generated from production titer response profiles obtained from a calibration set of clones with known fed batch production titer, and is configured to output the predicted relative fed batch production titer of the panel of clonal cells. 1. A rapid , high-throughput , computer-implemented method of predicting relative production titer of a panel of clonal producer cells in fed batch culture , the method comprising the steps of:{'sub': '50', 'simultaneously incubating each producer cell with at least three individual chemical cell stressors in static microplate culture for an incubation period of less than four days, in which each producer cell is incubated with a single chemical cell stressor provided at a concentration of 0.5 to 2.0 IC;'}after the incubation period determining the production titer responses of each cell in the presence and absence of the at least three chemical cell stressors to generate a cell-specific production titer response profile for each of the panel of producer cells; andinputting the cell-specific production titer response profile for each of the panel of ...

Bispecific anti-her2 antibody

The present invention relates to humanized bispecific anti-HER2 antibodies that comprise one antigen binding site containing variable regions of heavy and light chain of trastuzumab, and another antigen binding site containing variable regions of heavy and light chain of pertuzumab. The bispecific anti-HER2 antibodies is effective for treating cancer, such as breast cancer, gastric cancer, or ovarian cancer. Preferred bispecific anti-HER antibodies of the present invention are afucosylated antibodies. The present invention also relates to Chinese Hamster ovary (CHO) mutant cell line that has a dysfunctional Slc35C1 gene, which is the only dysfunctional gene in the mutant that affects glycan regulation.

ANIMAL PROTEIN-FREE MEDIA FOR CULTIVATION OF CELLS

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes. 1. An animal protein-free cell culture medium , comprising at least one polyamine and at least one protein hydrolysate derived from the group consisting of plants and yeast , wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to about 30 mg/L.2. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is selected from the group consisting of cadaverine claim 1 , putrescine claim 1 , spermidine claim 1 , spermine claim 1 , agmatine claim 1 , ornithine claim 1 , and a combination thereof.3. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is putrescine in a concentration ranging from about 0.5 to about 10 mg/L claim 1 , and the protein hydrolysate is soy hydrolysate in a concentration ranging from about 0.05% (w/v) to about 5% (w/v)4. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine originates from a source other than a protein hydrolysate.5. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to about 10 mg/L.6. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 2 to about 8 mg/L.7. The animal protein-free cell culture medium according to claim 1 , wherein the protein hydrolysate is present in ...

Methods and Compositions for the Clinical Derivation of an Allogenic Cell and Therapeutic Uses

Various cells, stem cells, and stem cell components, including associated methods of generating and using such cells are provided. In one aspect, for example, an isolated cell that is capable of self-renewal and culture expansion and is obtained from a subepithelial layer of a mammalian umbilical cord tissue. Such an isolated cell expresses at least three cell markers selected from CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and does not express at least three cell markers selected from CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR. 1. A method of treatment , comprising;administering isolated cells to a subject to treat a medical condition selected from COPD, diabetes, ischemia, osteoarthritis, orthopedic damage, liver damage, chronic refractory angina, erectile dysfunction, herniated disks, congestive heart failure, asthma, emphysema, wounds, acute radiation syndrome, autoimmune disorders, ischemic organ beds, graft vs. host disease, or a combination thereof, wherein the isolated cells are obtained, without enzymes, from a subepithelial layer of a mammalian umbilical cord tissue capable of self-renewal and culture expansion;wherein the isolated cell expresses at least three cell markers selected from the group consisting of CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105; andwherein the isolated cell does not express NANOG and at least five cell markers selected from the group consisting of CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.2. The method of claim 1 , wherein the isolated cell expresses CD29 claim 1 , CD73 claim 1 , CD90 claim 1 , CD166 claim 1 , SSEA4 claim 1 , CD9 claim 1 , CD44 claim 1 , CD146 claim 1 , and CD105.3. The method of claim 1 , wherein the isolated cell does not express CD45 claim 1 , CD34 claim 1 , CD14 claim 1 , CD79 claim 1 , CD106 claim 1 , CD86 claim 1 , CD80 claim 1 , CD19 claim 1 , CD117 claim 1 , Stro-1 claim 1 , and HLA-DR.4. The method of claim 1 , ...

Pd-1 antibodies

The present invention provides antibodies that bind human programmed cell death 1(PD-1), and may be useful for treating cancer alone and in combination with chemotherapy and other cancer therapeutics.

Mammalian Cell Culture-Produced Neublastin Antibodies

The present disclosure pertains to a mammalian cell culture genetically modified to express, and which expresses, a neublastin antibody polypeptide, or fragment thereof, in the culture, and to a neublastin antibody polypeptide, or fragment thereof, made by a mammalian cell culture genetically modified to express, and which expresses, the neublastin antibody polypeptide, or fragment thereof. 1. A cell culture comprising a mammalian cell line genetically modified to express , and which expresses , a neublastin antibody polypeptide , or a fragment thereof , in a cell culture medium.2. The cell culture of claim 1 , wherein the neublastin antibody polypeptide comprises SEQ ID NO:2 claim 1 , or a fragment thereof.3. The cell culture of claim 1 , wherein the neublastin antibody polypeptide comprises SEQ ID NO:4 claim 1 , or a fragment thereof.4. The cell culture of claim 3 , wherein the neublastin antibody polypeptide comprises SEQ ID NO:4 claim 3 , or a fragment thereof.5. The cell culture of claim 1 , wherein the mammalian cell line is a Chinese Hamster Ovary (CHO) cell line.6. The mammalian cell culture of claim 1 , wherein the cells have been adapted to grow in a serum-free medium claim 1 , an animal protein-free medium claim 1 , or a chemically defined medium.7. The cell culture of claim 1 , wherein the mammalian cells have been genetically modified with a polynucleotide encoding the neublastin antibody polypeptide claim 1 , or a fragment thereof.8. The mammalian cell culture of claim 1 , wherein the culture is a perfusion culture or is a fed batch culture.9. The mammalian cell culture of claim 1 , wherein the medium is a serum-free medium claim 1 , AN animal protein-free medium claim 1 , or a chemically defined medium.10. A neublastin antibody polypeptide produced in a large-scale mammalian cell culture claim 1 , the culture comprising mammalian cells genetically modified to express claim 1 , and which express claim 1 , the neublastin antibody polypeptide claim 1 , ...

LINKED PERFUSION TO CONTINUOUS-FLOW STIRRED-TANK REACTOR CELL CULTURE SYSTEM

Methods of protein production in a linked culture and production bioreactor system are provided. Such methods include a culture bioreactor (N-1 bioreactor) linked to production bioreactor (N bioreactor). More specifically, the methods include (a) culturing cells with a gene that encodes the protein of interest in a continuous perfusion culture bioreactor (N-1 bioreactor); inoculating a continuously stirred tank reactor (CSTR) production bioreactor (N bioreactor) with cells obtained from step (a); and culturing the cells in the CSTR production bioreactor under conditions that allow production of the protein of interest. 1. A method of producing a protein of interest , comprising:(a) culturing cells comprising a gene that encodes the protein of interest in a culture bioreactor (N-1 bioreactor);(b) inoculating a production bioreactor (N bioreactor) with cells obtained from step (a); and(c) culturing the cells in the production bioreactor under conditions that allow production of the protein of interest.2. The method according to claim 1 , wherein the method further comprises step (d) harvesting the protein of interest from the production bioreactor.3. The method according to claim 1 , wherein the culture bioreactor is a continuous perfusion culture bioreactor and the production bioreactor is a continuously stirred tank reactor (CSTR) production bioreactor.4. The method according to claim 1 , wherein the production bioreactor has no cell retention device.5. The method according to claim 1 , wherein volume ratio of the culture bioreactor to the production bioreactor is about 1:1 to about 1:20 or about 1:1 to about 1:5 or about 1:5.6. The method according to claim 1 , wherein the inoculation in step (b) is by transferring cells from the culture bioreactor to the production bioreactor.7. The method according to claim 6 , wherein the cell transfer is by cell bleed in continuous or semi-continuous modes.8. The method according to claim 7 , wherein the cell transfer is in ...

METHODS AND COMPOSITIONS FOR PRODUCTION OF FALLOPIAN TUBE EPITHELIUM

The fallopian tube epithelium (FTE) has been recognized as a site of origin of high-grade serous ovarian cancer (HGSC). However, absence of relevant in vitro human models that can recapitulate tissue-specific architecture has hindered understanding of FTE transformation and initiation of HGSC. Here, induced pluripotent stem cells (iPSCs) were used to establish a novel 3-dimensional (3D) human FTE organoid in vitro model containing the relevant cell types of the human fallopian tube as well as a luminal architecture that closely reflects the organization of fallopian tissues in vivo. Modulation of Wnt and nodal/activin signaling pathways provided iPSC differentiation into Müllerian cells and subsequent use of pro-Müllerian growth factors promoted FTE precursors. The expression of Müllerian markers verified correct cellular differentiation. An innovative 3D growth platform, which enabled the FTE organoid to self-organize into a convoluted luminal structure, permitted final differentiation to a FTE lineage. This powerful human-derived FTE organoid model can be used to study the earliest stages of HGSC development and to identify novel and specific biomarkers of early fallopian tube epithelial cell transformation. 1. A method for generating a fallopian tube epithelium (FTE) , comprising:providing a quantity of human pluripotent stem cells (hPSCs);culturing the hPSCs in the presence of at least one first growth factor and at least one induction molecule to generate mesoderm cells;further culturing the mesoderm cells in the presence of at least one second growth factor, at least one second induction molecule, and at least first one kinase inhibitor to generate intermediate mesoderm (IM) cells;additionally culturing the IM cells in the presence of at least one third growth factor, and at least one second kinase inhibitor to generate Mullerian epithelium cells; anddifferentiating Mullerian epithelium cells by addition of at least one fourth growth factor into FTE.2. The ...

System and Substances for Cryopreservation of Viable Cells

An improved cryopreservation process and substances can involve a cellular collection () in a cryopreservation fluid () that has been conditioned or treated () to enhance the cryopreservation process by adding () energy () such as in the surface energy of a substance in the cryopreservation fluid () prior to reducing energy for that same cryopreservation media for freezing. This can offer enhanced-post-cryogenic viability of the cryopreserved structures or a more optimum cooling curve () for a specific cell type.

FERMENTATION SYSTEMS

The present disclosure pertains to novel cell cultivation and cell and/or cell-derived product production processes that have advantages over currently existing fermentation strategies. The processes and methods according to the present disclosure may be used for an efficient supply of highly viable and metabolically active eukaryotic cells for transient production platforms, as an alternative production process with advantages over currently applied processes (batch, fed-batch or perfusion strategies) and for generating metabolically highly active biomass for subsequent use for transient expression systems or infection by a virus or pseudovirus or in cell-free systems. 1. A cell-density regulated cell cultivation process for the production of eukaryotic cells and/or an eukaryotic cell-derived product , wherein the cell density during the cell cultivation is regulated by adjusting the volume of the cell culture comprising the cells and a nutrient medium in a cultivation vessel , said process comprises the following steps:a) growing the cells in a cell culture having a variable cell culture volume, wherein the culture volume is not regulated to be constant over the entire cell cultivation process,b) measuring the cell density in the cell culture with a density sensor,c) regulating the cell density in the cell culture by adding an appropriate volume of the nutrient medium to the cell culture to keep the cells in the growth phase,d) harvesting a fraction of the cell culture at a desired cell density, wherein the harvested fraction comprises cells and/or a cell-derived product, and wherein the cell culture volume in the vessel is not kept constant continuously by adding nutrient medium into the vessel after harvesting said fraction and/or wherein the volume of the harvested fraction is not immediately replenished by adding nutrient medium into the vessel;e) repeating one or all of the aforementioned steps in the order set forth to allow a repeated harvest of cell ...

CONTRACEPTIVE VACCINES FOR MAMMALS

The invention provides a contraceptive vaccine for a female recipient of a target species of mammals including a composition of at least one of a plurality of granulosa cells and a plurality of ovarian stromal cells in combination with an adjuvant; wherein the cells are grown from a tissue sample of the cells obtained from at least one female donor of the target species which is not the same individual as the female recipient. The invention also provides a method of producing the contraceptive vaccine as well as a method of treating a female recipient with the contraceptive vaccine to prevent pregnancy. 1. A contraceptive vaccine for a female recipient of a target species of mammals comprising:a composition including at least one of a plurality of granulosa cells and a plurality of ovarian stromal cells in combination with an adjuvant;wherein the cells are grown from a tissue sample of the cells obtained from at least one female donor of the target species which is not the same individual as the female recipient.2. The contraceptive vaccine of wherein the target species is selected from at least one of the order of Primata claim 1 , Carnivora claim 1 , Diprotodontia claim 1 , Lagomorpha claim 1 , and Rodentia.3CanisFelis.. The contraceptive vaccine of claim 2 , wherein the target species selected from the order of Carnivora is selected from of at least one of the genus and the genus4. The contraceptive vaccine of claim 2 , wherein the target species selected from the order of Diprotodontia is selected from the family Didelphidae.5. The contraceptive vaccine of claim 2 , wherein the target species selected from the order of Lagomorpha is selected from the family Leporidae.6Rattus.. The contraceptive vaccine of claim 2 , wherein the target species selected from the order of Rodentia is selected from the genus7Homo sapiens.. The contraceptive vaccine of claim 2 , wherein the target species selected from the order of Primata is the species of8. The contraceptive vaccine ...

CELL LINE ADAPTED TO A PROTEIN-FREE AND LIPID-FREE MEDIUM, A METHOD FOR PRODUCING THE CELL LINE, AND A MEDIUM FOR THE CELL LINE

A cell of a cell line adapted to a protein-free and lipid-free medium, which is derived from CHO cells, can be stably used for production of recombinant proteins and can proliferate in a suspended state in a protein-free and lipid-free medium containing no exogenous growth factors. A method for adapting CHO cells by using a protein-free and lipid-free medium and a medium used for the method. 1. A cell of a cell line derived from Chinese Hamster Ovary (CHO) cells , the cell line being adapted to a protein-free and lipid-free medium , characterized in that the cell can proliferate in a suspended state in a protein-free and lipid-free medium comprising no exogenous growth factors.2. The cell as described in claim 1 , wherein the cell line has been deposited under Accession number NITE P-01641.3. A protein-free and lipid-free medium for culturing cells of an established cell line derived from CHO cells claim 1 , the cell line being adapted to a protein-free and lipid-free medium claim 1 , characterized by comprising putrescine claim 1 , thymidine claim 1 , hypoxanthine claim 1 , and monoethanolamine in a DMEM medium that has been modified so as to contain glucose in an amount of 3 to 5 times of the usual amount claim 1 , and by comprising no exogenous growth factors.4. The protein-free and lipid-free medium as described in claim 3 , which comprises 2000 to 5000 mg/L of glucose claim 3 , 0.001 to 2 mg/L of putrescine claim 3 , 0.01 to 1 mg/L of thymidine claim 3 , 0.1 to 10 mg/L of hypoxanthine claim 3 , and 0.1 to 5 mg/L of monoethanolamine.5. The protein-free and lipid-free medium as described in claim 3 , which further comprises 1 to 20 mg/L of insulin.6. A composition for producing the medium as described in claim 3 , which comprises claim 3 , in addition to the composition of the DMEM medium claim 3 , components of: 2000 to 5000 mg/L of glucose claim 3 , 0.001 to 2 mg/L of putrescine claim 3 , 0.01 to 1 mg/L of thymidine claim 3 , 0.1 to 10 mg/L of hypoxanthine ...

METHODS OF SELECTING AND DESIGNING SAFER AND MORE EFFECTIVE ANTI-CTLA-4 ANTIBODIES FOR CANCER THERAPY

The present invention relates to compositions of anti-CTLA-4 antibodies that bind to the human CTLA4 molecule and their use in cancer immunotherapy and for the reduction of autoimmune side effects compared to other immunotherapeutic agents. 1. An anti-CTLA-4 antibody for use in treating cancer , wherein the antibody does not confer systemic T cell activation or preferential expansion of self-reactive T cells.2. An anti-CTLA-4 antibody for use in treating cancer , wherein the antibody allows CTLA-4 to cycle back to a cell surface.3. The anti-CTLA-4 antibody of claim 2 , wherein the antibody binds to CTLA-4 with a higher affinity at pH 7 as compared to pH 5.5.4. The anti-CTLA-4 antibody of claim 2 , wherein the antibody binds to CTLA-4 with a higher affinity at pH 7 as compared to pH 4.5.5. The anti-CTLA-4 antibody of any one of - claim 2 , wherein the antibody induces FcR-mediated T regulatory cell depletion in a tumor microenvironment.6. The anti-CTLA-4 antibody of any one of - claim 2 , wherein the antibody does not confer systemic T cell activation or preferential expansion of self-reactive T cells.7. The anti-CTLA-4 antibody of any of the preceding claims claim 2 , wherein the antibody does not block binding of CTLA-4 to its B7 ligand.8. The anti-CTLA-4 antibody of any one of the preceding claims claim 2 , wherein the anti-CTLA-4 antibody has reduced affinity to soluble CTLA-4 compared to CTLA-4 located on the cell surface.9. The anti-CTLA-4 antibody of any of the preceding claims claim 2 , wherein the anti-CTLA-4 antibody is combined with an anti-PD-1 antibody or anti-PD-L1 antibody.11. The method of claim 10 , wherein the control anti-CTLA-4 antibody is Ipilimumab or Tremelimumab.12. The method of claim 10 , wherein the cells of step (a) express human CTLA-4.13. The method of claim 10 , wherein the cell surface CTLA-4 is detectably labeled.14. The method of claim 13 , wherein the detectable label is a fluorescent tag.15. The method of claim 14 , wherein the ...

REDUCTION OF HIGH MOLECULAR WEIGHT SPECIES, ACIDIC CHARGE SPECIES AND FRAGMENTS IN A MONOCLONAL ANTIBODY COMPOSITION

Alterations in bioreactor cell culture feeding to an extended or continuous feed following an initial period of no feeding reduces the level of high molecular weight, acid charge, and fragment species of monoclonal antibodies expressed in the culture, and enhances the level of afucosylated species of monoclonal antibodies expressed in the culture. Regular fucose infusions following an initial period of no feed media infusion reduces the level of afucosylated species of monoclonal antibodies expressed in the culture. Cell culture manipulation may be used to modulate the level of species of monoclonal antibodies. 1. A method for reducing one or more of high molecular weight species , acidic charge species , and fragments of a monoclonal antibody recombinantly expressed in a bioreactor , comprisingculturing recombinant cells that express the monoclonal antibody in a bioreactor, andbeginning on the second, third, fourth, or fifth day of the culture, and continuing every day until the conclusion of the culture, infusing the culture with a feed media continuously over a twenty four hour period sufficiently to reduce one or more of the high molecular weight species, acidic charge species, and fragments of the monoclonal antibody.2. The method according to claim 1 , wherein the monoclonal antibody specifically binds to tumor necrosis factor (TNF) alpha.3. The method according to any one of to claim 1 , wherein the recombinant cells comprise mammalian cells.4. The method according to any one of to claim 1 , wherein the recombinant cells comprise Chinese Hamster Ovary cells.5. The method according to any one of to claim 1 , wherein the recombinant cells comprise HEK293 cells.6. The method according to any one of to claim 1 , wherein the recombinant cells comprise Sp2/0 cells.7. The method according to any one of to claim 1 , wherein the high molecular weight species of the monoclonal antibody are reduced to about 5% or less of the total amount of monoclonal antibody expressed ...

CELL CULTURE MEDIA AND METHODS OF ANTIBODY PRODUCTION

Cell culture media are provided herein as are methods of using the media for cell culture and antibody production from cells. Compositions comprising antibodies and fragments thereof, produced by the methods herein are also provided. 1. A method of producing bevacizumab , or a fragment thereof , comprising the step of culturing a Chinese hamster ovary (CHO) cell comprising a nucleic acid encoding bevacizumab or fragment thereof in a cell culture medium , wherein the cell culture medium comprises two or more components selected from the group consisting of copper , insulin , and cystine , wherein the cell culture medium further comprises a plant-derived hydrolysate and an animal-derived hydrolysate , and wherein the cell produces bevacizumab , or a fragment thereof.2. The method of claim 1 , wherein the cell culture medium comprises copper and insulin.3. The method of claim 1 , wherein the cell culture medium comprises copper and cystine.4. The method of claim 1 , wherein the cell culture medium comprises insulin and cystine.5. The method of claim 1 , wherein the cell culture medium comprises copper claim 1 , insulin claim 1 , and cystine.6. (canceled)7. The method of claim 1 , wherein the cell culture medium comprises insulin at a concentration of from about 1.0 mg/L to about 100.0 mg/L.8. The method of claim 1 , wherein the cell culture medium comprises insulin at a concentration of from about 10.0 mg/L to about 100.0 mg/L.9. The method of claim 1 , wherein the cell culture medium comprises insulin at a concentration of from about 10.0 mg/L to about 50.0 mg/L.10. The method of claim 1 , wherein the cell culture medium comprises insulin at a concentration of from about 10.0 mg/L to about 35.0 mg/L.11. The method of claim 1 , wherein the cell culture medium comprises insulin at a concentration of from about 10.0 mg/L to about 25.0 mg/L.12. The method of claim 1 , wherein the cell culture medium comprises insulin at a concentration of about 25 mg/L.13. The method of ...

CELL CULTURE MEDIUM FOR ADAMTS PROTEIN EXPRESSION

The present invention provides culture mediums that are useful for the expression of ADAMTS proteins, such as ADAMTS13. Methods for the expression and purification of ADAMTS proteins are also provided. In some embodiments, the mediums and methods of the invention are useful for the expression of ADAMTS proteins having high specific activities. Also provided are ADAMTS, e.g., ADAMTS13, protein compositions with high specific activities, which are expressed and purified according to the methods provided herein. 121-. (canceled)22. A method for expressing a disintegrin and metalloproteinase with thrombospondin motifs 13 (ADAMTS13) protein , the method comprising culturing , by continuous cultivation , a mammalian cell harboring a nucleic acid encoding an ADAMTS13 protein in a culture medium comprising at least 3 μM zinc and at least 4 mg/L nicotinamide.23. The method of claim 22 , wherein the culture medium comprises from 3 μM to 12 μM zinc.24. The method of claim 22 , wherein the culture medium comprises about 5 μM zinc.25. The method of claim 22 , wherein the culture medium comprises from 4 mg/L to 10 mg/L nicotinamide.26. The method of claim 23 , wherein the culture medium comprises from 4 mg/L to 10 mg/L nicotinamide.27. The method of claim 24 , wherein the culture medium comprises from 4 mg/L to 10 mg/L nicotinamide.28. The method of claim 22 , wherein the culture medium comprises about 7 mg/L nicotinamide.29. The method of claim 23 , wherein the culture medium comprises about 7 mg/L nicotinamide.30. The method of claim 24 , wherein the culture medium comprises about 7 mg/L nicotinamide.31. The method of claim 22 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.32. The method of claim 26 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.33. The method of claim 30 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.34. The method of claim 22 , wherein the mammalian cell is selected from ...

High purity ovarian cancer stem cells for active autologous immune therapy

The disclosure provides cancer stem cells, for use in stimulating immune response against a cancer, such as ovarian carcinoma. Methods for preparing and purifying the cancer stem cells are provided.

Methods for determining developmental stage of human cumulus cells

The present invention relates generally to the fields of reproductive medicine. More specifically, the present invention relates to a method determining the developmental stage of human cumulus cells issues from MII oocyte.

3D MICROPHYSIOLOGIC SYSTEM

The present invention relates generally to a three-dimensional cell and tissue culture system for the female reproductive tract. In particular provided herein the system includes individual female reproductive cultures in a dynamic microfluidic setting or integrated using a microfluidic microphysiologic system. In some embodiments, the present invention provides ex-vivo female reproductive tract integration in a three dimensional (3D) microphysiologic system. 131.-. (canceled)32. A microphysiologic system comprising:(a) a first 3D cell culture subsystem comprising at least a first female reproductive cell type in 3D culture and a culture media for said first female reproductive cell type; and(b) a second 3D cell culture subsystem comprising at least a second female reproductive cell type in 3D culture and a culture media for said second female reproductive cell type;wherein the first 3D cell culture subsystem and the second 3D cell culture subsystem are in unidirectional fluid communication such that fluid from the first 3D cell culture subsystem flows downstream to the second 3D cell culture subsystem.33. The microphysiologic system of claim 32 , wherein the first and second 3D cell culture subsystems are selected from the group consisting of ovarian claim 32 , fallopian claim 32 , uterine claim 32 , endocervical claim 32 , and ectocervical subsystems.34. The microphysiologic system of claim 33 , wherein the first and second 3D cell culture subsystems are distinct types of culture subsystems.35. The microphysiologic system of claim 34 , wherein the first 3D cell culture subsystem is selected from the group consisting of ovarian claim 34 , fallopian claim 34 , uterine claim 34 , and endocervical subsystems.36. The microphysiologic system of claim 35 , wherein the second 3D cell culture subsystem is selected from the group consisting of fallopian claim 35 , uterine claim 35 , endocervical claim 35 , and ectocervical subsystems.37. The microphysiologic system of claim ...

ANTIBODIES WITH MODULATED GLYCAN PROFILES

This invention relates to recombinantly-expressed denosumab molecules and methods for modulating glycan profiles of denosumab molecules. 2. The method of claim 1 , wherein during the growth phase claim 1 , the glucose concentration is maintained at from 4 g/L to 20 g/L by bolus feed or perfusion.3. The method of claim 2 , wherein when the host cells are incubated in the second culture medium during the production phase claim 2 , the glucose concentration is maintained at from 0 g/L to 8 g/L claim 2 , and the galactose concentration is maintained at from 7 g/L to 15 g/L claim 2 , by bolus feed or perfusion.4. The method of claim 1 , wherein during the production phase claim 1 , the host cells are initially maintained in the first culture medium for about 3 to about 15 days claim 1 , and subsequently transitioned into the second culture medium by perfusion or bolus feed.5. The method of claim 1 , wherein in step (a) claim 1 , said cell density is from 5×10viable cells/mL to 12×10viable cells/mL.6. The method of claim 1 , wherein from about 4% to about 11% of the denosumab molecules comprise high-mannose at the N-298 site.7. The method of claim 1 , wherein said mammalian host cell is a CHO cell.8. The method of claim 1 , wherein said mammalian host cell is a CS-9 cell.9. The method of claim 1 , wherein said first culture medium comprises methotrexate (MTX).10. The method of claim 1 , comprising:(a) incubating said mammalian host cell in a first culture medium during growth phase, and supplementing the culture with one or more bolus feeds, wherein the glucose concentration is maintained at from about 4 g/L to about 18 g/L during the growth phase;(b) transitioning host cells from step (a) from growth phase to production phase, and maintaining the glucose concentration at from about 4 g/L to about 18 g/L for about 3 days to about 15 days; and subsequently(c) transitioning the host cells of (b) into a second culture medium, wherein said second culture medium comprises from ...

Methods of cell culture

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described.

CELL CULTURE MEDIUM, AND CELL CULTURE APPARATUS AND CELL CULTURE METHOD EACH USING SAME

A cell culture apparatus () of the present invention comprises: a culture vessel () in which a cell culture medium comprising at least one of culture components composed of a conjugate with a stimuli-responsive polymer is stored and cells are cultured in the medium; a stimulus-applying mechanism () that applies a predetermined stimulus to the conjugate so as to induce a predetermined change of the stimuli-responsive polymer in response to the stimulus; and, a separation mechanism () that separates at least a part of the medium components except for the conjugate from the cell culture medium, while leaving the conjugate in the cell culture medium, on the basis of a property change of the stimuli-responsive polymer. 1. A cell culture medium comprising at least one of medium components that is a conjugate with a stimuli-responsive polymer.2. The cell culture medium as claimed in claim 1 , wherein the stimuli-responsive polymer changes in response to at least one stimulus of temperature claim 1 , light and pH.3. The cell culture medium as claimed in claim 2 , wherein the stimuli-responsive polymer claim 2 , at least claim 2 , elongates its molecular chain or changes its polarization charge in response to the stimulus.4. The cell culture medium as claimed in claim 1 , wherein the stimuli-responsive polymer is at least one of polylysine claim 1 , polyglutamine and polyarginine.5. The cell culture medium as claimed in claim 1 , wherein the conjugate is composed of the stimuli-responsive polymer and a cell growth factor.6. The cell culture medium as claimed in claim 5 , wherein the cell growth factor is at least one growth factor selected from the group consisting of epidermal growth factor (EGF) claim 5 , insulin-like growth factor (IGF) claim 5 , transforming growth factors (TGF) claim 5 , nerve growth factor (NGF) claim 5 , brain-derived neurotrophic factor (BDNF) claim 5 , vesicular endothelial growth factor (VEGF) claim 5 , Glanulocitye-Colony Stimulating Factor (G-CSF ...

PREPARING ANTIBODIES FROM CHO CELL CULTURES FOR CONJUGATION

The invention is based in part on the observation that a CHO cell oxidizing enzyme, particularly QSOX1, can survive a seemingly rigorous antibody purification process to reduce subsequent conjugation efficiency of the antibody to a drug. Whether the oxidizing enzyme survives the purification procedure depends on which purification techniques are employed which can vary from one antibody to another. With knowledge that contamination with a CHO cell oxidizing enzyme is a potential problem for subsequent conjugation, a suitable purification scheme can be devised for any antibody that eliminates or at least reduces CHO oxidizing enzyme(s) to an acceptable level. 1. A method of producing a conjugated antibody , comprising:(a) obtaining a preparation of antibody;(b) testing the preparation for the presence of a CHO cell sulfhydryl oxidizing enzyme selected from at least one of quiescin Q6 sulfhydryl oxidase 1 (QSOX1), Q6 sulfhydryl oxidase 2 (QSOX2), and Augmenter of Liver Regeneration (ALR);(c) if the enzyme is detected at a detectable level in step (b), performing a purification step to remove the enzyme to a non-detectable level, wherein the purification step is selected from a step of (i) loading the preparation on a protein A column with a salt wash, (ii) performing depth filtration, (iii) using anion exchange with a quaternary ammonium ion column, and (iv) performing phenyl membrane filtration;(d) if the enzyme is not detected at a detectable level in step (b) or (c), proceeding to step (d); and(e) conjugating the at least partially purified antibody via one or more sulfhydryl groups to a drug to produce the conjugated antibody.2. The method of claim 1 , wherein the CHO cell sulfhydryl oxidizing enzyme is QSOX1.3. The method of claim 1 , wherein the testing comprises identifying a band of 65-75 kDa on a gel.4. The method of claim 3 , wherein the band is identified by western blot or silver stain.5. The method of claim 1 , wherein the testing comprises a functional ...

MEDIUM FOR THE PROTEIN-FREE AND SERUM-FREE CULTIVATION OF CELLS

A medium is described for the protein-free and serum-free cultivation of cells, especially mammalian cells, whereby the medium contains a proportion of soy hydrolysate. 1. A method of adapting recombinant cells to serum-free and animal protein-free medium , the method comprising:a. culturing recombinant cells in serum-containing medium,b. transferring said cells into a serum-free and animal protein-free medium, wherein the medium comprises soy hydrolysate wherein at least 40% of said soy hydrolysate has a molecular weight of ≦500 Daltons, andc. culturing said cells in said serum-free and animal protein-free medium.2. The method of claim 1 , wherein the medium contains a quantity in excess of 10 wt % soy hydrolysate based on the total dry weight of the medium.3. The method of claim 1 , wherein said medium contains ultrafiltered soy hydrolysate.4. The method of claim 3 , wherein the medium contains purified soy hydrolysate.5. The method of claim 1 , wherein the soy hydrolysate has an endotoxin content of <500 U/g.6. The method of claim 1 , wherein at least 500 of the soy hydrolysate has a molecular weight of ≦500 Daltons.7. The method of claim 6 , wherein at least 550 of the soy hydrolysate has a molecular weight of ≦500 Daltons.8. The method of claim 1 , wherein the medium also contains an amino acid.9. The method of claim 7 , wherein the amino acid is selected from the group consisting of L-asparagine claim 7 , L-cysteine claim 7 , L-cystine claim 7 , L-proline claim 7 , L-tryptophan claim 7 , and L-glutamine claim 7 , or mixtures thereof.10. The method of claim 1 , wherein the medium also contains one or more auxiliary substances selected from the group consisting of buffer substances claim 1 , oxidation stabilizers claim 1 , stabilizers to counteract mechanical stress claim 1 , and protease inhibitors.11. The method of claim 1 , wherein the cells contain a coding sequence for a recombinant blood factor claim 1 , and are capable of expressing the recombinant blood ...

COMPOSITIONS AND METHODS FOR CULTURING CELLS FROM NORMAL HUMAN TUBO-OVARIAN EPITHELIUM AND HUMAN TUBO-OVARIAN TUMORS

Described herein are cell culture media, kits and methods for preparing cell culture media, and methods for culturing cells, for example, cells of the female reproductive tract, and tumor cells. 1. A cell culture medium , or a kit for preparing a cell culture medium , comprising:(a) adenosine triphosphate;(b) a carrier protein;(c) cholesterol, linoleic acid, and lipoic acid;(d) glutathione;(e) a nucleotide salvage pathway precursor base selected from hypoxanthine, xanthine, adenine, guanine and thymidine;(f) phosphoethanolamine;(g) selenium;(h) transferrin;(i) triiodothyronine;(j) vitamin A, vitamin C, and vitamin D;(k) Zn, Mg, and Cu;(l) an agent that increases intracellular cAMP;(m) epidermal growth factor (EGF);(n) hydrocortisone;(o) insulin; and(p) serum.2. The cell culture medium of claim 1 , further comprising one or more of:(a) adenosine monophosphate;(b) vitamin E; or(c) at least one of vitamin K3, niacin, or niacinamide.3. (canceled)4. The cell culture medium of claim 1 , wherein the carrier protein is albumin.57-. (canceled)8. The cell culture medium of claim 1 , wherein the agent that increases intracellular cAMP is cholera toxin.924-. (canceled)25. The cell culture medium of claim 1 , further comprising an estrogen.2627-. (canceled)28. The cell culture medium of claim 25 , wherein the estrogen is 17-beta-estradiol.29. The cell culture medium of claim 1 , wherein the medium is substantially free of estrogen.3036-. (canceled)37. The cell culture medium of claim 1 , wherein the medium supports proliferation of ovarian tumor cells for at least about 15 population doublings (PD) in vitro.38. The cell culture medium of claim 1 , wherein the medium supports proliferation of ovarian cells and/or fallopian tube cells for at least about 15 population doublings (PD) in vitro.39. A kit for preparing the cell culture medium of claim 1 , comprising a first one or more containers comprising components (a)-(k) and a second one or more containers comprising components (l ...

CELL CULTURE MEDIUM FOR ADAMTS PROTEIN EXPRESSION

The present invention provides culture mediums that are useful for the expression of ADAMTS proteins, such as ADAMTS13. Methods for the expression and purification of ADAMTS proteins are also provided. In some embodiments, the mediums and methods of the invention are useful for the expression of ADAMTS proteins having high specific activities. Also provided are ADAMTS, e.g., ADAMTS13, protein compositions with high specific activities, which are expressed and purified according to the methods provided herein. 121-. (canceled)22. A method for expressing a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) protein , the method comprising culturing , by continuous cultivation , a mammalian cell harboring a nucleic acid encoding an ADAMTS protein in a culture medium comprising at least 3 μM zinc , wherein the continuous cultivation includes maintaining a cell density of from 1×10cells/mL to 4×10cells/mL for at least seven days.23. The method of claim 22 , wherein the culture medium comprises from 3 μM to 12 μM zinc.24. The method of claim 22 , wherein the culture medium comprises about 5 μM zinc.25. The method of claim 22 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.26. The method of claim 23 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.27. The method of claim 24 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.28. The method of claim 22 , wherein the mammalian cell is selected from the group consisting of a CHO cell claim 22 , a BHK cell claim 22 , and an HEK cell.29. The method of claim 23 , wherein the mammalian cell is selected from the group consisting of a CHO cell claim 23 , a BHK cell claim 23 , and an HEK cell.30. The method of claim 24 , wherein the mammalian cell is selected from the group consisting of a CHO cell claim 24 , a BHK cell claim 24 , and an HEK cell.31. The method of claim 22 , wherein the continuous cultivation is chemostatic cell ...

SERUM-FREE MAMMALIAN CELL CULTURE MEDIUM, AND USES THEREOF

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. 178-. (canceled)79. A method of producing a polypeptide comprising(a) obtaining a mammalian cell that has been genetically engineered to produce a polypeptide; and(b) cultivating said cell in a serum-free and protein-free cell culture medium comprising an insulin substitute and a transferrin substitute, under conditions favoring expression of said polypeptide by said mammalian cell.80. The method of claim 79 , wherein said mammalian cell is an epithelial cell.81. The method of claim 79 , wherein said mammalian cell is a human cell.82. The method of claim 81 , wherein said human cell is a 293 embryonic kidney epithelial cell.83. A polypeptide produced according to the method of .84. A eukaryotic cell culture medium comprising a Fe chelate and a Zn salt claim 79 , wherein said medium is capable of supporting the high-density growth of mammalian cells in suspension culture and/or the expression of recombinant protein.85. (canceled)86. The eukaryotic cell culture medium according to claim 84 , wherein said mammalian cells are Chinese hamster ovary cells.87. The eukaryotic cell culture medium according to claim 84 , wherein said ...

DECREASING ORNITHINE METABOLISM TO DECREASE THE HIGH MANNOSE GLYCOFORM CONTENT OF RECOMBINANT PROTEINS

The present invention relates to a method for manipulating the high mannose glycoform content of recombinant glycoproteins by regulating ornithine metabolism during cell culture. 156.-. (canceled)57. A method of increasing high mannose glycoform content of a recombinant protein comprising culturing a host cell expressing the recombinant protein in a cell culture comprising one selected from the group consisting of ornithine , arginine , an ornithine aminotransferase inhibitor , a nitric oxide synthase inhibitor , an ornithine decarboxylase inhibitor , and an arginine decarboxylase inhibitor , wherein ornithine production in the host cell is increased when compared to the host cell expressing the recombinant protein is cultured in a cell culture lacking one selected from the group consisting of ornithine , arginine , an ornithine aminotransferase inhibitor , a nitric oxide synthase inhibitor , ornithine decarboxylase inhibitor , and an arginine decarboxylase inhibitor , andthe recombinant protein has an increased high mannose glycoform content than when the recombinant protein is expressed in the cell culture lacking one selected from the group consisting of ornithine, arginine, an ornithine aminotransferase inhibitor, a nitric oxide synthase inhibitor, ornithine decarboxylase inhibitor, and an arginine decarboxylase inhibitor.58. The method of claim 57 , whereinthe ornithine aminotransferase inhibitor is 5-fluoromethylornithine;{'sup': 'G', 'the nitric oxide synthase inhibitor is selected from the group consisting of 2-ethyl-2-thiopseudourea and N-Nitro-L-arginine and L-monomethyl-L-arginine; or'}the arginine decarboxylase inhibitor is asymmetric dimethyl-arginine.59. The method of claim 57 , wherein ornithine accumulation in the host cell is increased by the addition of at least 0.6 mM ornithine to the cell culture.60. The method of claim 57 , wherein the concentration of ornithine is selected from the group consisting of 0.6 to 14.8 mM; 6 to 14.8 mM; 0.6 mM claim ...

Animal Protein-Free Media for Cultivation of Cells

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes. 1. A method for cultivating cells , comprising the steps of:(a) providing an animal protein-free cell culture medium;(b) adding a supplement to the animal protein-free cell culture medium comprising at least one polyamine and at least one protein hydrolysate derived from the group consisting of plants and yeast, wherein the resultant concentration of the at least one polyamine in the animal protein-free cell culture medium ranges from about 0.5 to about 30 mg/L; and(c) propagating the cells in the animal protein-free cell culture medium to form a cell culture.2. The method according to claim 1 , wherein the cells are selected from the group consisting of mammalian cells claim 1 , insect cells claim 1 , avian cells claim 1 , bacterial cells claim 1 , and yeast cells.3. The method according to claim 2 , wherein the mammalian cells are CHO cells.4. The method according to claim 3 , wherein the CHO cells are propagated in suspension.5. The method according to claim 4 , wherein the cell culture is a chemostat suspension culture.6. The method according to claim 1 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation claim 1 , feed-batch-cultivation claim 1 , perfusion cultivation claim 1 , and chemostat-cultivation.7. The method according to claim 1 , wherein the polyamine is selected from the group consisting of cadaverine claim 1 , putrescine claim 1 , spermidine claim 1 , spermine claim 1 , agmatine claim 1 , ornithine ...

Method for increasing the specific production rate of eukaryotic cells

The current invention reports the use of meta-tyrosine for increasing the specific productivity of a eukaryotic cell that produces/expresses a polypeptide. In the current method it is not necessary to perform a temperature-, osmolality- or pH shift or to add drugs like valproic acid or sodium butyrate to modulate the specific productivity of the cultivated cells. The method does not affect cell viability or product titer. 1. A method for increasing specific productivity (qP) of a Chinese Hamster Ovary (CHO) cell , the method comprising culturing the cell in culture medium , wherein the culture medium contains meta-tyrosine.29-. (canceled)10. The method of claim 1 , wherein the culture medium further comprises phenylalanine in a non-limiting concentration.1115-. (canceled)16. A method for producing an exogenous polypeptide in a Chinese Hamster Ovary (CHO) cell claim 1 , wherein the CHO cell expresses a nucleic acid encoding the exogenous polypeptide claim 1 , the method comprising culturing the CHO cell in a culture medium comprising meta-tyrosine.17. The method of claim 16 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.2 mM to 0.7 mM.18. The method of claim 16 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.25 mM to 0.6 mM.19. The method of claim 16 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.5 mM.20. The method of claim 16 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.4 mM.2122-. (canceled)23. The method of claim 16 , wherein the polypeptide is an immunoglobulin or a variant thereof or a fragment thereof or a fusion thereof.24. The method of claim 16 , wherein the culture medium additionally comprises phenylalanine in a non-limiting concentration.25. The method of claim 24 , wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 1.25.26. The method of claim 24 , wherein the molar ...

MICROCARRIER PERFUSION CULTURING METHODS AND USES THEREOF

Provided herein are methods of culturing a mammalian cell and various methods that utilize these culturing methods. 1110.-. (canceled)111. A method for testing a manufacturing process for making a recombinant protein , the method comprising:providing a shake tube containing an adherent Chinese hamster ovary (CHO) cell containing a nucleic acid encoding a recombinant protein disposed in a first liquid culture medium, wherein the first liquid culture medium occupies about 10% to about 30% of the volume of the shake tube and contains a plurality of microcarriers at a concentration of about 1.0 g/L to about 15.0 g/L;incubating the shake tube for a period of time at about 32° C. to about 39° C. and with a rotary agitation of about 130 revolutions per minute (RPM) to about 150 RPM;{'sup': '6', 'after about the first 48 to 96 hours of the period of time, continuously or periodically removing a first volume of the first liquid culture medium and adding to the first liquid culture medium a second volume of a second liquid culture medium, wherein the first and second volumes are about equal, and the method achieves a viable cell density of greater than 2.0×10cells/mL in the first liquid culture medium or a combination of the first and second liquid culture medium at some point during the period of time;'}detecting the recombinant protein in the cell or in the first and/or second culture medium; andcomparing the amount of recombinant protein present in the adherent CHO cell or in the first and/or second culture medium to a reference level of recombinant protein.112. The method of claim 111 , wherein the reference level of recombinant protein is a level of recombinant protein produced using a different culturing method.113. The method of claim 112 , wherein the different culturing method utilizes a different first or second liquid culture medium claim 112 , a different adherent CHO cell claim 112 , a different temperature claim 112 , a different level of agitation claim 112 , a ...

SYNTHETIC HYDROGELS FOR ORGANOGENESIS

Synthetic hydrogels for organogenesis support organogenesis from mammalian cells, including human cells. The synthetic hydrogels typically include a network of crosslinked branched biodegradable polymers. A portion of the branches of the branched biodegradable polymers are linked to binders which are generally synthetic peptides for cell and extracellular matrix attachment. The hydrogels may include an inhibitor of apoptosis. The synthetic hydrogels with the synthetic binders typically do not interfere with cellular, proteomic, genetic, and/or transcriptome analyses of organoids formed in the hydrogel. The synthetic hydrogels may be subject to on-demand dissolution to provide intact organoids substantially free of hydrogel polymers. Also provided are methods of making the synthetic hydrogels and methods of using the synthetic hydrogels for organogenesis. 1. A synthetic hydrogel cell culture matrix comprising:biodegradable polymers andbinders comprising one or more adhesion ligands and one or more peptide binders at a molar ratio between about 1.2:1 and 5:1, and, optionally,one or more inhibitors of apoptosis.2. The synthetic hydrogel of claim 1 , wherein the biodegradable polymers are branched biodegradable polymers.3. The synthetic hydrogel of claim 1 , wherein the biodegradable polymers comprise one or more polyalkylene glycols claim 1 , preferably polyethylene glycols.4. The synthetic hydrogel of claim 1 , wherein the biodegradable polymers have molecular weight between about 2 kDa and about 100 kDa.5. The synthetic hydrogel of claim 1 , wherein the biodegradable polymers are functionalized biodegradable polymers.6. The synthetic hydrogel of claim 1 , wherein the binders are synthetic peptides.7. The synthetic hydrogel of claim 1 , wherein the binders comprise one or more adhesion ligands and one or more peptide binders at a molar ratio between about 2:1 and about 5:1.8. The synthetic hydrogel of claim 1 , wherein the one or more adhesion ligands are cell-binding ...

METABOLICALLY OPTIMIZED CELL CULTURE

An improved method for large scale production of proteins and/or polypeptides in cell culture is provided. In accordance with the present invention, the method provides for culturing cells that have metabolically shifted. The use of such a method or system allows high levels of protein or polypeptide production and reduces accumulation of unwanted metabolic waste such as lactate. Proteins and polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical, immunogenic, or other commercial biologic compositions, such as antibodies. 118.-. (canceled)19. A method for culturing cells comprising:(a) culturing cells in a first cell culture, wherein the first cell culture is a seed train cell culture, wherein lactate accumulates in the first cell culture, and wherein said lactate in the first cell culture consists of lactate produced by the cells in the first cell culture;(b) determining a metabolic shift to lactate consumption has occurred in the first cell culture, wherein the metabolic shift to lactate consumption results from accumulation of lactate produced in the first cell culture; and(c) transferring the cells from the first cell culture to a second cell culture after the metabolic shift to lactate consumption in the cells of the first cell culture has occurred, wherein the second cell culture is a production culture, and wherein said transferring cells to the second cell culture comprises transferring cells from the first cell culture to a production bioreactor.20. The method of claim 19 , wherein the cells are transfected with DNA encoding a polypeptide of interest prior to culturing cells in the first cell culture claim 19 , and comprising maintaining the second cell culture under conditions that allow the expression of the polypeptide of interest claim 19 , and harvesting the polypeptide of interest from the second cell culture.21. The method of claim 20 , wherein the polypeptide of interest is ...

Nucleic acid constructs for va rna transcription

Herein is reported a novel adenoviral VA RNA nucleic acid wherein the wild-type type 2 polymerase III promoter has been removed and an U6-snRNA promoter or an inducible promoter has been added.

Cell culture media and methods of antibody production

Cell culture media are provided herein as are methods of using the media for cell culture and antibody production from cells. Compositions comprising antibodies and fragments thereof, produced by the methods herein are also provided.

ANIMAL PROTEIN-FREE MEDIA FOR CULTIVATION OF CELLS

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes. 1. A method for cultivating Chinese hamerster ovary (CHO) cells in soy hydrolysate-containing animal protein-free medium supplemented with purescine to overcome inhibitory effects on cell growth due to soy hydrolysate lot variation , said method comprising the steps of:(a) providing an animal protein-free cell culture medium comprising soy hydrolysate and putrescine, wherein the putrescine is added to said culture medium in a concentration ranging from 0.5 mg/L to 10 mg/L, and the soy hydrolysate is present in a concentration ranging from 0.05% (w/v) to 0.5% (w/v); and(b) propagating the cells in the medium to form a cell culture.2. The method according to claim 1 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation claim 1 , feed-batch-cultivation claim 1 , perfusion cultivation claim 1 , and chemostat-cultivation. This application is a continuation of U.S. patent application Ser. No. 13/864,118 filed Apr. 16, 2013, which is a continuation of U.S. patent application Ser. No. 12/965,111 filed Dec. 10, 2010, issued U.S. Pat. No. 8,440,408; which is a continuation of U.S. patent application Ser. No. 11/858,844, filed Sep. 20, 2007, now abandoned; which is a division of U.S. patent application Ser. No. 10/976,399, filed Oct. 29, 2004, now abandoned; each of which applications is herein incorporated by reference in its entirety for all purposes.The present invention relates to animal protein-free cell culture media ...

Compositions and Methods for Enhancing Bioenergetic Status in Female Germ Cells

Compositions and methods comprising bioenergetic agents for restoring the quality of aged oocytes, enhancing oogonial stem cells or improving derivatives thereof (e.g., cytoplasm or isolated mitochondria) for use in fertility-enhancing procedures, are described. 1. A composition comprising a mammalian oocyte , a mammalian oogonial stem cell (OSC) , the progeny of a mammalian OSC or a mammalian preimplantation zygote , a CD38 inhibitor and a nicotinamide adenine dinucleotide (NAD precursor) , wherein:the OSC is obtained from ovarian tissue, is an isolated non-embryonic stem cell, and is mitotically competent and expresses Vasa, Oct-4, Dazl and Stella and, optionally, a stage-specific embryonic antigen;the CD38 inhibitor is a compound selected from the group consisting of apigenin, luteolin, tyrphostin-8, berberine and SRT-1720; andthe NAD precursor is a compound selected from the group consisting of nicotinamide mononucleotide, nicotinamide riboside or nicotinic acid; andwherein the CD38 inhibitor and the NAD precursor are present in the composition in an amount effective to enhance the bioenergetic status of said oocyte, OSC, progeny of an OSC or preimplantation zygote in said composition.2. The composition of claim 1 , further comprising a culture medium selected from the group consisting of cell culture medium claim 1 , oocyte retrieval solution claim 1 , oocyte washing solution claim 1 , oocyte in vitro maturation medium claim 1 , ovarian follicle in vitro maturation medium claim 1 , oocyte in vitro fertilization medium claim 1 , vitrification solution and cryopreservation solution.3. The composition of claim 1 , wherein the NAD precursor is nicotinamide riboside.4. The composition of claim 1 , wherein the NAD precursor is nicotinic acid.5. The composition of claim 1 , wherein a mammalian OSC is cultured.6. The composition of claim 1 , wherein the mammalian oocyte claim 1 , or OSC claim 1 , progeny of an OSC or preimplantation zygote is from a human female.7. The ...

SCAFFOLDS FOR UTERINE CELL GROWTH

A method for growing polarized endometrial cells, said method comprising: (a) disposing endometrial cells on a scaffold, said scaffold comprising a silica-based glass composition, characterized by multi-modal porosity, said scaffold being to define a top side and a bottom side; (b) providing nutrients to said top and bottom sides of said scaffold and an environment to grow polarized endometrial cells on said scaffold. 1. A method for growing polarized endometrial cells , said method comprising:disposing endometrial cells on a scaffold, said scaffold comprising a silica-based glass composition, characterized by multi-modal porosity, said scaffold being to define a top side and a bottom side;providing nutrients to said top and bottom sides of said scaffold and an environment to grow polarized endometrial cells on said scaffold.2. The method of claim 1 , wherein the porosity is characterized by multi-modal porosity.3. The method of claim 2 , wherein the bi-modal porosity comprises interconnected microstructure of nanopores and macropores.4. The method of claim 3 , wherein the macropores have an average pore diameter of greater than about 10 micrometers.5. The method of claim 3 , wherein the nanopores have an average diameter of less than about 100 nanometers.6. The method of claim 1 , wherein the composition comprises a glass of the formula: 30% CaO-70% SiO.7. The method of claim 6 , wherein the composition comprises a glass of the formula: 24.5 CaO-(27.5−x)NaO-6PO-(42+x)SiO(wt %) claim 6 , wherein x is from about 0 to about 10.8. The method of claim 7 , wherein x is about 3.9. The method of claim 1 , wherein the composition comprises a glass of the formula: 24.4% NaO-26.9% CaO-2.6% PO-46.1% SiO(mol %).10. The method of claim 1 , wherein said scaffold is substantially planar.11. The method of claim 1 , wherein said scaffold is substantially flat.12. The method of claim 1 , wherein said scaffold is produced using melt-quench method of fabrication.13. The method of claim ...

METHOD OF CELL CULTURE

A method of cell culture comprising providing cells in a cell culture medium to start a cell culture process, and, 1. A method of cell culture comprising(i) providing cells in a cell culture medium to start a cell culture process, and,(ii) maintaining at least one metabolite selected from 3-(4-hydroxyphenyl)lactate, 4-hydroxyphenylpyruvate, phenyllactate, indolelactate, indolecarboxylic acid, homocysteine, 2-hydroxybutyric acid, isovalerate and formate below a concentration C1 in the cell culture medium, wherein C1 is 3 mM.2. The method of wherein C1 is 1 mM.3. The method of wherein step ii) comprises the step of measuring the concentration of said at least one metabolite claim 1 , and claim 1 , when the measured concentration is above a predefined value claim 1 , the concentration of precursor of said at least one metabolite in the cell culture medium is decreased by reducing the amount of precursor provided to the cells.4. The method of wherein said concentration of said at least one metabolite is measured online using NMR claim 3 , HPLC or UPLC.5. The method of wherein claim 3 ,when the measured concentration of 3-(4-hydroxyphenyl)lactate, 4-hydroxyphenylpyruvate and/or phenyllactate is above said predefined value, the concentration of phenylalanine is decreased in the cell culture medium and/or,when the measured concentration of 3-(4-hydroxyphenyl)lactate and/or 4-hydroxyphenylpyruvate is above said predefined value, the concentration of tyrosine is decreased in the cell culture medium, and/or,when the measured concentration of indolelactate and/or indolecarboxylic acid is above said predefined value, the concentration of tryptophan is decreased in the cell culture medium and/or,when the measured concentration of homocysteine and/or 2-hydroxybutyric acid is above said predefined value, the concentration of methionine is decreased in the cell culture medium, and/or,when the measured concentration of isovalerate is above said predefined value, the concentration of ...

CELL CULTURE PROCESS FOR PRODUCING A PROTEIN

The invention provides the mammalian cell culture process for the production of monoclonal antibodies and fusion proteins wherein the mammalian cell is cultured in suitable cell culture conditions specifically maintaining monophasic temperature. 1. A process of producing glycosylated protein in a mammalian cell culture the process comprising the steps of:a) preparing inoculum with suitable cell concentration during seed development,b) inoculating the inoculum with suitable cell concentration in to production bioreactor,c) culturing the cell in production bioreactor at suitable conditions wherein the suitable condition is monophasic temperature condition; andd) obtaining the glycosylated protein from the cell culture.2. The process as claimed in claim 1 , wherein monophasic temperature is selected from the range of about 32° C. to about 37° C.3. The process as claimed in claim 1 , wherein monophasic temperature is selected from the range of about 34° C. to about 37° C.4. The process as claimed in claim 2 , wherein monophasic temperature is about 33° C.5. The process as claimed in claim 2 , wherein monophasic temperature is about 34° C.6. The process as claimed in claim 2 , wherein monophasic temperature is about 35° C.7. The process as claimed in claim 2 , wherein monophasic temperature is about 36° C.8. The process as claimed in claim 1 , wherein monophasic temperature does not comprise temperature shift.9. The process as claimed in claim 1 , wherein the mammalian cell is selected from CHO cell.10. The process as claimed in claim 9 , wherein the CHO cell is a dhfr-CHO cell.11. The process as claimed in claim 1 , wherein the mammalian cells are cultured in fed-batch mode in production bioreactor.12. The process as claimed in claim 1 , wherein the cell culture process does not have distinctive growth phase and production phase.13. The process as claimed in claim 1 , wherein the suitable conditions further comprises pH selected from 6.7 to 7.4.14. The process as ...

CELL SORTING METHOD AND SYSTEM

A cell sorting method includes: obtaining a cervical sample of a pregnant mammal, the cervical sample including placental trophoblast cells and cervical cells; removing the mucus of the cervical sample; dispersing the placental trophoblast cells and the cervical cells; centrifuging the cervical sample to remove the supernatant of the cervical sample; and using a dielectrophoretic chip to perform sorting on the cervical sample, so as to sort out the placental trophoblast cells from the cervical cells. 1. A cell sorting method , comprising:obtaining a cervical sample of a pregnant mammal, the cervical sample including placental trophoblast cells and cervical cells;removing mucus in the cervical sample;dispersing the placental trophoblast cells and the cervical cells;centrifuging the cervical sample to remove supernatant in the cervical sample; andusing a dielectrophoretic chip to perform sorting on the cervical sample, so as to sort out the placental trophoblast cells from the cervical cells.2. The cell sorting method of claim 1 , wherein the pregnant mammal is a pregnant woman.3. The cell sorting method of claim 1 , wherein the cervical sample is collected at which a pregnancy of the pregnant mammal is 5week to 20week.4. The cell sorting method of claim 1 , wherein using the dielectrophoretic chip to sort out the placental trophoblast cells and the cervical cells is performed in an environment of about 4° C.5. The cell sorting method of claim 1 , further comprising:fixing the cervical sample by using a reservoir.6. The cell sorting method of claim 1 , further comprising:{'sup': 5', '5, 'solving the cervical sample in a conductive solution after removing supernatant in the cervical sample, such that a cell density of the cervical sample achieves about 2×10cells/ml to 5×10cells/ml and that a conductivity of the cervical sample achieves less than 50 μS/cm.'}7. The cell sorting method of claim 6 , wherein the conductive solution includes 0.25-0.5% bovine serum albumin ( ...

PH ADJUSTMENT TO IMPROVE THAW RECOVERY OF CELL BANKS

Provided herein are methods of freezing mammalian cells for storage or improving thaw recovery of cell banks comprising freezing mammalian cells in a freezing medium having a pH of 6.7 to 8.5. 1. A method of improving thaw recovery of cell banks comprising freezing mammalian cells for banking in a freezing medium , wherein the freezing medium comprises a buffered solution and a cryoprotective agent , and wherein the freezing medium has a pH of about 6.7 to about 8.5 prior to freezing.2. A method of freezing mammalian cells for storage comprising freezing the mammalian ceils in a freezing medium , wherein the freezing medium comprises a buffered solution and a cryoprotective agent , and wherein the freezing medium has a pH of about 6.7 to about 8.5 prior to freezing.3. The method of or , wherein the freezing medium has a pH of about 6.7 to about 8.3 , about 7.2 to about 7.8 , or about 7.5 prior to freezing.4. The method of or , wherein the pH of the freezing medium has been adjusted to a pH of about 6.7 to about 8.5.5. The method of or , wherein the pH of the freezing medium has been adjusted to a pH of about 6.7 to about 8.3 or about 7.2 to about 7.8.6. The method of or , wherein the pH of the freezing medium has been adjusted to a pH of about 7.5.7. The method of any one of - , wherein the cells are combined with a freezing medium before and/or after pH adjustment.8. The method of any one of - , wherein the adjusted pH is a target pH or a measured pH.9. The method of claim 8 , wherein the target pH is about 6.7 to about 8.3.10. The method of claim 8 , wherein the measured pH is about 6.7 to about 8.3.11. The method of any one of - claim 8 , further comprising a step of measuring an initial pH of the freezing medium containing the cells prior to adjusting pH of the freezing medium.12. The method of any one of - claim 8 , further comprising a step of measuring the adjusted pH of the freezing medium.13. The method of claim 12 , wherein if the measured pH of the ...

METHODS OF INCREASING PROTEIN PRODUCTION IN MAMMALIAN CELLS

Aspects of the present disclosure provide compositions and methods for increasing protein production in mammalian cells, e.g. methods of increasing mammalian cell expression of a protein of interest, comprising culturing mammalian cells that overexpress a protein of interest and are modified to overexpress a gene encoding Rab 11 or Yap1, as well as mammalian cells that overexpress a protein of interest and which are modified to overexpress a gene encoding Rab 11 or Yap1. 1. A method of increasing mammalian cell expression of a protein of interest , comprising culturing mammalian cells that overexpress a protein of interest and are modified to overexpress a gene encoding Rab11 protein.2. The method of claim 1 , wherein the cells are cultured in cell culture media under conditions that permit production and secretion of the protein of interest into the media.3. The method of claim 2 , further comprising isolating and/or purifying the protein of interest from the media.4. The method of any one of - claim 2 , wherein the mammalian cells comprise a recombinant nucleic acid encoding the Rab11 protein and/or a recombinant nucleic acid encoding the protein of interest.5. The method of any one of - claim 2 , wherein the Rab11 protein is stably expressed in the mammalian cells.6. The method of any one of - claim 2 , wherein the protein of interest is stably expressed in the mammalian cells.7. The method of any one of - claim 2 , wherein the mammalian cells are Chinese hamster ovary (CHO) cells.8. The method of any one of - claim 2 , wherein the Rab11 protein is a Rab11a isoform or a Rab11b isoform.9. The method of claim 8 , wherein the Rab11 protein is a Rab11b isoform.10. The method of any one of - claim 8 , wherein the protein of interest is a therapeutic protein.11. The method of claim 10 , wherein the therapeutic protein is an antibody.12. The method of claim 11 , wherein the antibody is a monoclonal antibody.13. The method of any one of - claim 11 , wherein the cell ...

CELL CULTURE PROCESS

A cell culture medium comprising tyrosine at a concentration of at least 3 mM and polyvinylalcohol (PVA). 1. A cell culture medium comprising tyrosine at a concentration of at least 4 mM and polyvinylalcohol (PVA).2. (canceled)3. The cell culture medium according to wherein the concentration of tyrosine is at least 10 mM.4. The cell culture medium according to wherein the concentration of tyrosine is between 5 and 20 mM.5. The cell culture medium according to wherein the concentration of PVA is at least 0.5 g/L.6. The cell culture medium according to wherein the concentration of PVA is between 0.5 and 5 g/L.7. (canceled)8. (canceled)9. The cell culture medium according to wherein said medium is protein free.10. The cell culture medium according to wherein the turbidity is less than 5NTU after two weeks storage at 4° C. in the absence of light.11. (canceled)12. The cell culture medium according to wherein the medium comprises 4 to 10 mM Ala claim 1 , 30 to 60 mM Arg claim 1 , 50 to 90 mM Asn claim 1 , 10 to 30 mM Asp claim 1 , 2 to 40 mM Glu claim 1 , 2 to 15 mM Gly claim 1 , 8 to 20 mM His claim 1 , 25 to 32 mM Ile claim 1 , 35 to 60 mM Leu claim 1 , 28 to 60 mM Lys claim 1 , 9 to 25 mM Met claim 1 , 10 to 30 mM Phe claim 1 , 15 to 40 mM Pro claim 1 , 44 to 80 mM Ser claim 1 , 20 to 45 mM Thr claim 1 , 2 to 10 mM Trp and 20 to 50 mM Val.13. A method of cell culture comprising contacting mammalian cells with a cell culture medium according to .14. (canceled)15. The method of claim 13 , wherein the mammalian cells are CHO cells.16. (canceled)17. The method of claim 13 , wherein the cell culture is a fed batch culture.18. The method of claim 17 , wherein the fed batch culture comprises a base medium supplemented with feed media.19. The method of claim 17 , wherein only the base medium is a medium according to .20. The method of wherein only the feed medium is a medium according to .21. The method of claim 17 , wherein the base medium and the feed medium are media ...

INFORMATION PROCESSING APPARATUS, INFORMATION PROCESSING SYSTEM, INFORMATION PROCESSING METHOD, AND PROGRAM

[Object] To provide an information processing apparatus, an information processing system, an information processing method, and a program which are suitable for assessing a fertile ovum with high precision. 1. An information processing apparatus , comprising:a storage unit that pre-stores a plurality of first images, the plurality of first images being obtained by rotating and imaging a comparative cell;an image acquisition unit that acquires a plurality of second images, the plurality of second images being obtained by rotating and imaging a cell that is an object to be assessed; andan assessment unit that assesses the cell that is the object to be assessed on a basis of a result of comparison of the first image with the second image.2. The information processing apparatus according to claim 1 , whereinthe assessment unit assesses the cell that is the object to be assessed on a basis of a result of comparison of a first feature quantity extracted from the first image with a second feature quantity extracted from the second image.3. The information processing apparatus according to claim 2 , whereinthe storage unit stores an assessment result of the first image which is associated with the first image, further comprising:a determination unit that compares the first image with the second image and determines whether or not the first image is identical to the second image; anda giving unit that gives the second image the assessment result associated with the first image determined to be identical to the second image in a case where the determination unit determines that the first image is identical to the second image, whereinthe assessment unit considers a most given assessment result of assessment results respectively given to the plurality of second images as an assessment of the cell that is the object to be assessed.4. The information processing apparatus according to claim 3 , further comprising:a calculation unit that calculates an occupancy rate of the most ...

METHODS FOR IN VITRO MATURATION OF OVARIAN FOLLICLES

The invention relates to methods to be used in the maturation of ovarian follicles and oocytes. More specifically, the invention concerns the use of inhibitors of the phosphatase PTEN, such as oxovanadate and peroxovanadate complexes, in methods for in vitro and in vivo maturation of follicles and oocytes. 1. (canceled)2. A method for the in vitro activation of mammalian non-growing follicles , the method comprising the step of incubating the mammalian follicles in a physiological acceptable medium comprising one or more inhibitors of PTEN.3. The method according to claim 2 , wherein the non-growing follicles claim 2 , are selected from primordial claim 2 , intermediate and primary follicles.4. The method according to claim 2 , wherein the activated follicles are further used for in vitro fertilization.5. The method according to claim 2 , wherein the non-growing follicles are human.7. A method for the in vitro maturation of an oocyte which comprises the activation of non-growing follicles according to .8. A method of in vitro fertilization comprising implanting an embryo claim 7 , wherein said embryo is produced by a method comprising treating a mature oocyte with sperm claim 7 , wherein said oocyte is produced by the method of . The invention relates to methods to be used in in vitro maturation of ovarian follicles and oocytes. More specifically the invention concerns the use of inhibitors of the phosphatase PTEN in methods for in vitro maturation of follicles and oocytes.Inducing activation of ovarian follicles in order to achieve maturation is highly desirably from a research perspective but also within numerous fields of application. Primordial follicles could potentially serve as a source of oocytes for in vitro fertilization, but the ability to utilize activated and matured ovarian follicles for other applications, for instance post-chemotherapy or radiation treatment of cancer, is also of great importance. However, there is currently no means for primordial ...

MAMMALIAN CELL CULTURE PROCESSES FOR PROTEIN PRODUCTION

The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, preferably, but not limited to, fed-batch cell cultures. In one aspect, the methods comprise the addition of glucocorticoid compound during the culturing period. The addition of glucocorticoid compound sustain a high viability of the cultured cells, and can yield an increased end titer of protein product, and a high quality of protein product, as determined, e.g., by sialic acid content of the produced protein. 1. A method of increasing sialylation of a recombinant CTLA4 molecule , comprising: a) culturing CHO cells which produce a recombinant CTLA4 molecule in cell culture under conditions that allow for protein production; and b) feeding the CHO cells with feeding medium containing dexamethasone , wherein the recombinant CTLA4 molecule comprises the extracellular domain of CTLA4 with the amino acid sequence beginning with methionine at position 27 or alanine at position 26 and ending at aspartic acid at position 150 as shown in SEQ ID NO: 2 joined to an immunoglobulin moiety comprising hinge , CH2 and CH3 domains and wherein the dexamethasone is sustained or maintained in the CHO cell culture at a concentration of 0.001 μM to 10 μM , wherein the CHO cell culture volume is at least 500 liters , and wherein sialylation of the recombinant CTLA4 molecule is increased compared to sialylation recombinant CTLA4 molecule in a culture without dexamethasone addition.2. A method of reducing cell death rate in a CHO cell culture producing recombinant CTLA4 molecule , comprising: a) culturing CHO cells which produce a recombinant CTLA4 molecule in cell culture under conditions that allow for protein production; and b) feeding the CHO cells with feeding medium containing dexamethasone , wherein the recombinant CTLA4 molecule comprises the extracellular domain of CTLA4 with the amino acid sequence beginning with methionine at ...

Mammalian cell culture processes for protein production

The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, preferably, but not limited to, fed-batch cell cultures. In one aspect, the methods comprise the addition of glucocorticoid compound during the culturing period. The addition of glucocorticoid compound sustain a high viability of the cultured cells, and can yield an increased end titer of protein product, and a high quality of protein product, as determined, e.g., by sialic acid content of the produced protein.

METHOD FOR CULTURING MULTIPLE OVARIAN FOLLICLES BY USING ANGIOTENSIN II RECEPTOR AGONIST

A method for culturing multiple ovarian follicles by using an angiotensin II receptor agonist, including a first culturing step of culturing a cluster of separated multiple follicles in medium, and a second culturing step of further culturing the cluster of multiple follicles cultured in the culturing step, in medium containing an angiotensin II receptor agonist, and to a composition for culturing a cluster of multiple follicles. The composition contains an angiotensin II receptor agonist. 1. A follicle culturing method comprising:(a) a first culturing step of culturing a cluster of separated multiple follicles in medium; and(b) a second culturing step of further culturing the cluster of multiple follicles cultured in the first culturing step, in medium containing an angiotensin II receptor agonist.2. The follicle culturing method of claim 1 , wherein the cluster of separated multiple follicles is in a pre-antral stage.3. The follicle culturing method of claim 1 , wherein the cluster of multiple follicles includes two to four pre-antral follicles joined together.4. The follicle culturing method of claim 1 , wherein the angiotensin II receptor agonist is one selected from a group consisting of angiotensin II claim 1 , CGP-42112 and Novokinin trifluoroacetate salt.5. The follicle culturing method of claim 4 , wherein a concentration of the CGP-42112 is in a range of 5 ug/mL to 20 ug/mL in the medium.6. The follicle culturing method of claim 1 , wherein the first culturing step is performed for 2 days to 6 days.7. The follicle culturing method of claim 1 , wherein the second culturing step is performed for 5 days to 20 days.8. The follicle culturing method of claim 1 , further including a step of obtaining an ovum by inducing ovulation of the cluster of multiple follicles cultured in the second culturing step.9. A composition for culturing a cluster of multiple follicles claim 1 , wherein the composition contains an angiotensin II receptor agonist.10. The composition ...

METHOD FOR PRODUCING ARTIFICIAL SYNCYTIOTROPHOBLAST AND PROGENITOR CELL THEREOF

According to the present invention, there is provided a method for producing artificial trophoblasts derived from human cells, that includes the steps of: adhesion culturing human pluripotent stem cells in a culture medium containing a BMP signal transduction activator, and during the culturing bringing the cells under culturing into contact with at least one selected from the group consisting of a γ aminobutyric acid B receptor activator, a peroxisome proliferator-activated receptor γ activator, a retinoid X receptor activator, and a retinoic acid receptor activator, thereby obtaining a culture containing trophoblasts differentiated from the human pluripotent stem cells, and so on. 1. A method for producing artificial trophoblasts derived from human cells , comprising the steps of: adhesion culturing human pluripotent stem cells in a culture medium containing a BMP signal transduction activator , and during the culturing bringing the cells under culturing into contact with at least one selected from the group consisting of a γ aminobutyric acid B receptor activator , a peroxisome proliferator-activated receptor γ activator , a retinoid X receptor activator , and a retinoic acid receptor activator , thereby obtaining a culture containing trophoblasts differentiated from the human pluripotent stem cells.2. A method for producing artificial syncytiotrophoblasts derived from human cells , comprising the steps of: adhesion culturing human pluripotent stem cells in a culture medium containing a BMP signal transduction activator , and during the culturing bringing the cells under culturing into contact with at least one selected from the group consisting of a γ aminobutyric acid B receptor activator , a peroxisome proliferator-activated receptor γ activator , a retinoid X receptor activator , and a retinoic acid receptor activator , thereby obtaining a culture containing syncytiotrophoblasts differentiated from the human pluripotent stem cells.3. The production method ...

Anti-VHH Domain Antibodies and Use Thereof

The invention provides a group of anti-VHH domain antibodies and use thereof. The present invention further provides use of the described antibodies in the development, screening and purification of nano-antibodies, and use of the described antibodies in the field of immunotherapy. 1. (canceled)2. An antibody or an antigen-binding fragment thereof , wherein the antibody contains a heavy chain variable region and a light chain variable region , wherein (A) the heavy chain variable region comprises heavy chain complementarity determining region (HCDR) 1 , HCDR2 and HCDR3 , wherein (a) the HCDR1 has a sequence selected from an amino acid sequence shown in SEQ ID NO: 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 or 15 or an amino acid sequence having at least 90% identity thereto , (b) the HCDR2 has a sequence selected from an amino acid sequence shown in SEQ ID NO: 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 or 30 or an amino acid sequence having at least 90% identity thereto , and (c) the HCDR3 has a sequence selected from an amino acid sequence shown in SEQ ID NO: 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 or 45 or an amino acid sequence having at least 90% identity thereto; and (B) the light chain variable region comprises light chain complementarity determining region (LCDR) 1 , LCDR2 and LCDR3 , wherein (d) the LCDR1 has a sequence selected from an amino acid sequence shown in SEQ ID NO: 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 or 60 or an amino acid sequence having at least 90% identity thereto , (e) the LCDR2 has a sequence selected from an amino acid sequence shown in SEQ ID NO: 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 or 75 or an amino acid sequence having at least 90% identity thereto , and (f) the LCDR3 has a sequence selected from an amino acid sequence shown in SEQ ID NO: 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 or ...

MESENCHYMAL STEM CELL THERAPY FOR SPINAL MUSCULAR ATROPHY

Disclosed are means, methods and compositions of matter useful for treatment of spinal muscular atrophy. In one embodiment, stem cells of the mesenchymal type are modified to enhance anti-inflammatory and regenerative potential in a manner to prevent disease, inhibit progression and/or reverse existing disease. In other embodiments combinations of mesenchymal stem cells together with extracts and/or products derived from said mesenchymal stem cells are administered for prevention, inhibition of progression and/or reversion of spinal muscular atrophy. 1. A method of ameliorating the effects of spinal muscular atrophy comprising the steps of: a) identifying a subject suffering spinal muscular atrophy; b) providing a population of stem cells , and/or derivatives of stem cells; and b) administering said stem cells and/or derivatives of stem cells to said subject at a concentration and frequency sufficient to ameliorate the effects of spinal muscular atrophy.2. The method of claim 1 , wherein said ameliorated effects of spinal muscular atrophy are selected from the group consisting of: muscle strengthening claim 1 , improved balance claim 1 , improved fine motor skills claim 1 , lessened tremors claim 1 , improved appetite claim 1 , improved ability to eat claim 1 , improvement in walking.3. The method of claim 2 , wherein said spinal muscular atrophy is caused by mutations in the Survival Motor Neuron (SMN) gene.4. The method of claim 3 , wherein said mutations of said SMN gene is associated with reduction in SMN1 protein.5. The method of claim 1 , wherein said spinal muscular atrophy is selected from a group consisting of: a) Type 1 spinal muscular atrophy; b) Type 2 spinal muscular atrophy; c) Type 3 spinal muscular atrophy; and d) Type 4 spinal muscular atrophy.6. The method of claim 1 , wherein said stem cells are mesenchymal stem cells.7. The method of claim 6 , wherein said mesenchymal stem cells are plastic adherent.8. The method of claim 6 , wherein said ...

ENGINEERED MAMMALIAN CELLS FOR PRODUCTION OF RECOMBINANT PROTEINS

The present invention relates to mammalian cells modified to provide for improved expression of a recombinant protein of interest. In particular, the invention relates to CHO cells and other host cells in which the expression of one or more endogenous secreted proteins has been disrupted, as well as to the preparation, identification and use of such cells in the production of recombinant proteins. 1. A CHO cell modified to disrupt expression of one or more endogenous proteins selected from the group consisting of TIMP1 , NID1-1 , NID1-2 , BGN , and LGALS3BP.2. The CHO cell according to claim 1 , wherein the CHO cell is modified to disrupt expression of TIMP1.3. The CHO cell according to claim 1 , wherein the CHO cells is modified to disrupt expression of LGALS3BP.4. The CHO cell according to claim 1 , wherein the CHO cell is modified to disrupt expression of NID1-1 claim 1 , NID1-2 claim 1 , or both.5. The CHO cell according to claim 1 , wherein the CHO cell is modified to disrupt expression ofat least 5 endogenous proteins;at most 50 endogenous proteins; orbetween 5 and 50 endogenous proteins.6. The CHO cell according to claim 1 , wherein the CHO cell is modified to disrupt expression of TIMP1 claim 1 , LGALS3BP claim 1 , NID1-1 claim 1 , NID1-2 claim 1 , BGN and CTSD.7. The CHO cell according to claim 1 , wherein the expression of at least one endogenous protein is disrupted by knockdown of the gene encoding the endogenous protein.8. The CHO cell according to claim 1 , wherein the expression of at least one endogenous protein is disrupted by knockout of the gene encoding the endogenous protein.9. The CHO cell according to claim 1 , which comprises a nucleic acid sequence encoding a recombinant protein of interest claim 1 , the nucleic acid sequence being extrachromosomal or chromosomally integrated and under the control of an inducible or constitutive promoter.10. The CHO cell according to claim 9 , which cell produce higher levels of said recombinant protein of ...

USE OF N-ACETYL-5-METHOXYTRYPTAMINE OR ANALOGUES THEREOF, FOR PROMOTING THE MECHANISM OF IMPLANTATION OF THE EMBRYO AND RELATED COMPOSITIONS AND CULTURE MEDIA

The present invention refers to the use of N-acetyl-5-methoxytryptamine (melatonin) and/or an analogue thereof, for use in the medical or veterinary field in the assisted reproduction for promoting the mechanism of implantation of the embryo, and in particular for the prevention of implantation failure into the uterus, by topical administration of an effective amount in a mammalian subject female in need of such treatment, and related compositions, culture media and medical devices. 1. N-acetyl-5-methoxy tryptamine or an analogue thereof , for use in the medical or veterinary field in assisted reproduction for the prevention of implantation failure into the uterus by topical administration of an effective amount in a mammalian subject female in need of such treatment.2. N-acetyl-5-methoxy tryptamine or an analogue thereof claim 1 , according to claim 1 , wherein said analogue is selected from agomelatine claim 1 , 6-hydroxymelatonin claim 1 , serotonin claim 1 , 5 hydroxytryptophan or their derivatives.32. N-acetyl-5-methoxy tryptamine or an analogue thereof claims 1 , according to any one of - claims 1 , wherein said female mammalian subject is a woman suffering from infertility or polyabortion.43. N-acetyl-5-methoxy tryptamine or an analogue thereof claims 1 , according to any one of - claims 1 , wherein said topical administration is via endometrial irrigation or uterine washing or endometrial washing.54. N-acetyl-5-methoxy tryptamine or an analogue thereof claims 1 , according to any one of - claims 1 , wherein said topical administration is carried out in a single administration at the time of oocyte retrival.65. N-acetyl-5-methoxy tryptamine or an analogue thereof claims 1 , according to any one of - claims 1 , wherein said N-acetyl-5-methoxy tryptamine or analogue thereof is present in a concentration ranging from 4×10g/ml to 25×10g/ml claims 1 , preferably greater than or equal to 10×10g/ml.76. N-acetyl-5-methoxy tryptamine or an analogue thereof claims 1 , ...

MEDIUM COMPOSITION FOR CULTURING ANIMAL CELLS FOR PRODUCING RECOMBINANT EXTRACELLULAR MATRIX PROTEIN AND METHOD OF USING THE SAME

Provided are a medium composition for culturing animal cells for producing a recombinant extracellular matrix protein, a method of producing the recombinant extracellular matrix protein with high purity, and a method of assaying a monomer of the recombinant extracellular matrix protein. 1. A method of assaying a monomer of a recombinant extracellular matrix protein , the method comprising:performing size exclusion chromatography on a sample comprising the recombinant extracellular matrix protein using a mobile phase comprising hydrochloride; andanalyzing the monomer of the recombinant extracellular matrix protein in the sample, based on a result of the size exclusion chromatography.2. The method of claim 1 , wherein the recombinant extracellular matrix protein is collagen claim 1 , elastin claim 1 , fibronectin claim 1 , laminin claim 1 , vitronectin claim 1 , tenascin claim 1 , or hyaluronan and proteoglycan link (HAPLN) protein.3. The method of claim 2 , wherein the recombinant HAPLN protein is any one protein selected from the group consisting of HAPLN1 claim 2 , HAPLN2 claim 2 , HAPLN3 claim 2 , and HAPLN4.4. The method of claim 1 , wherein the hydrochloride is arginine hydrochloride claim 1 , aniline hydrochloride claim 1 , adenine hydrochloride claim 1 , guanine hydrochloride claim 1 , guanidine hydrochloride claim 1 , histidine hydrochloride claim 1 , or lysine hydrochloride.5. The method of claim 1 , wherein the mobile phase comprises hydrochloride at a concentration of more than 0.5 M.6. The method of claim 1 , wherein claim 1 , in the analyzing claim 1 , ratios of the monomer of the recombinant extracellular matrix protein and other impurities are analyzed. This is a divisional of U.S. patent application Ser. No. 17/781,266, filed May 31, 2022, which is the § 371 U.S. National Stage of International Application No. PCT/KR2021/020184, filed Dec. 29, 2021, which in turn claims the benefit of Korean Patent Application No. 10-2020-0188062, filed on Dec. 30, ...

Method for increasing the specific production rate of eukaryotic cells

The current invention reports the use of meta-tyrosine for increasing the specific productivity of a eukaryotic cell that produces/expresses a polypeptide. In the current method it is not necessary to perform a temperature-, osmolality- or pH shift or to add drugs like valproic acid or sodium butyrate to modulate the specific productivity of the cultivated cells. The method does not affect cell viability or product titer. 1. A method for increasing specific productivity (qP) of a eukaryotic host cell , the method comprising culturing the cell in culture medium , wherein the culture medium contains meta-tyrosine.2. The method of claim 1 , wherein the eukaryotic host cell is a mammalian cell.3. The method of claim 2 , wherein the mammalian cell is a Chinese Hampster Ovary (CHO) cell.4. The method of claim 1 , wherein the eukaryotic host cell produces an exogenous polypeptide.5. The method of claim 1 , wherein the eukaryotic host cell is cultured in suspension.6. The method of claim 1 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.2 mM to 0.7 mM.7. The method of claim 1 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.25 mM to 0.6 mM.8. The method of claim 1 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.5 mM.9. The method of claim 1 , wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.4 mM.10. The method of claim 1 , wherein the culture medium further comprises phenylalanine in a non-limiting concentration.11. The method of claim 10 , wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 1.25.12. The method of claim 10 , wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.25.13. The method of claim 10 , wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.125.14. The method of claim 10 , wherein the molar ratio of meta-tyrosine/ ...

MANGANESE SUPPLEMENTATION FOR CONTROL OF GLYCOSYLATION IN MAMMALIAN CELL CULTURE PROCESS

The present invention pertains to a cell culture medium comprising manganese as a media supplement, which was shown to control recombinant protein glycosylation and methods of using thereof. The present invention further pertains to a method of controlling or manipulating glycosylation of a recombinant protein of interest in a large scale cell culture. 1. A method for achieving a predetermined glycosylation profile of an anti-α4-integrin antibody comprising providing manganese to a cell culture at a concentration that falls within a target manganese concentration range , wherein the cell culture comprises host cells producing the anti-α4-integrin antibody.2. The method of claim 1 , comprising supplementing the cell culture with manganese if the manganese concentration in the cell culture is below the target manganese concentration range.3. A method for achieving a predetermined glycosylation profile of an anti-α4-integrin antibody comprising (i) determining a manganese concentration in a component of a cell culture medium claim 1 , (ii) if the manganese concentration is below a target manganese concentration range claim 1 , supplementing the cell culture medium with the component to achieve a manganese concentration within the target manganese concentration range claim 1 , and (iii) culturing a recombinant host cell producing an anti-α4-integrin antibody in the cell culture medium comprising the cell culture medium component.4. A method for achieving a predetermined glycosylation profile of an anti-α4-integrin antibody comprising (i) determining a manganese concentration in a component of a cell culture medium claim 1 , (ii) if the manganese concentration is below a target manganese concentration range claim 1 , adding manganese to the component of the cell culture medium to achieve a manganese concentration within the target manganese concentration range claim 1 , (iii) producing a cell culture medium using the component of cell culture medium with the target ...

METHODS FOR MODULATING PROTEIN GALACTOSYLATION PROFILES OF RECOMBINANT PROTEINS USING PERACETYL GALACTOSE

The present invention relates to methods and compositions for modulating glycosylation profile, such as the galactosylation profile, of recombinant proteins expressed by mammalian host cells during the cell culture process by supplementing cell culture media with a peracetyl galactose. 115-. (canceled)16. A method of producing a recombinant protein with a modulated galactosylation profile , said method comprising culturing a host cell expressing said protein in: a) cell culture medium comprising a peracetyl galactose; or b) cell culture medium complemented with at least one feed comprising a peracetyl galactose.17. The method according to claim 16 , further comprising purifying said recombinant protein with a modulated galactosylation profile.18. The method according to claim 16 , wherein the modulation of the galactosylation level is a decrease in the galactosylation level in said protein.19. The method according to claim 16 , wherein the peracetyl galactose is α-2-F peracetyl galactose or β-2-F peracetyl galactose.20. The method according to claim 16 , wherein the host cell is Chinese Hamster Ovary (CHO) cells.21. The method according to claim 16 , wherein the recombinant protein is selected from the group consisting of an antibody or antigen binding fragment thereof claim 16 , a human antibody or antigen-binding portion thereof claim 16 , a humanized antibody or antigen-binding portion thereof claim 16 , a chimeric antibody or antigen-binding portion thereof claim 16 , a recombinant fusion protein claim 16 , a growth factor claim 16 , a hormone claim 16 , or a cytokine.22. The method according to claim 16 , wherein the concentration of peracetyl galactose in the cell culture medium before seeding is of or of about 0.1 μM to 200 μM.23. The method according to claim 16 , wherein the concentration of peracetyl galactose in the cell culture medium after seeding is of or of about 0.08 μM to 180 μM.24. A composition comprising a cell culture medium comprising a ...

PREPARING ANTIBODIES FROM CHO CELL CULTURES FOR CONJUGATION

The invention is based in part on the observation that a CHO cell oxidizing enzyme, particularly QSOX1, can survive a seemingly rigorous antibody purification process to reduce subsequent conjugation efficiency of the antibody to a drug. Whether the oxidizing enzyme survives the purification procedure depends on which purification techniques are employed which can vary from one antibody to another. With knowledge that contamination with a CHO cell oxidizing enzyme is a potential problem for subsequent conjugation, a suitable purification scheme can be devised for any antibody that eliminates or at least reduces CHO oxidizing enzyme(s) to an acceptable level. 1. A method of producing a conjugated antibody , comprising:(a) performing at least one purification step of a purification scheme to obtain at least a partially purified preparation of an antibody from a culture of CHO cells expressing the antibody;(b) testing the preparation for presence of a CHO cell oxidizing enzyme;(c) if an unacceptable level of the enzyme is detected as present in the preparation of step (b), repeating steps (a) and (b) with a different purification step;(d) if an acceptable level or no CHO cell oxidizing enzyme is detected as present in the preparation of step (b), performing at least one purification step resulting in the acceptable level or no detected CHO cell oxidizing enzyme on the same culture or a second culture of CHO cells expressing the antibody to obtain at least a partially purified preparation of antibody.2. The method of claim 1 , further comprising step (e) conjugating the at least partially purified antibody via one or more sulfhydryl groups to a drug to produce the conjugated antibody.3. The method of claim 1 , wherein the CHO cell oxidizing enzyme is QSOX1.4. The method of wherein the second culture in step (d) is a larger culture by volume than the culture in step (a).5. The method of claim 4 , wherein the second culture in step (d) is at least 1000 times larger by ...

METHODS OF MAKING ALKALINE PHOSPHATASE AGENTS

The present invention relates to, inter alia, compositions and methods, including therapeutic alkaline phosphatases that find use in the treatment of disease, such as microbiome-related diseases. In part, the invention provides, in various embodiments, methods of manufacturing therapeutic alkaline phosphatases. 1. A method of producing a recombinant IAP comprising:a) providing an initial culture of mammalian cells, the mammalian cells comprising a gene encoding said IAP;b) batch feeding said culture;c) adding a quantity of supplemental zinc to said culture; andd) isolating the produced IAP from said mammalian cells.2. The method of claim 1 , wherein said supplemental zinc is selected from the group consisting of ZnSO claim 1 , ZnCl claim 1 , ZnBr claim 1 , zinc citrate claim 1 , hydrolysate claim 1 , and plasma zinc bound to serum albumin.3. The method of claim 2 , wherein said batch feeding occurs during the culturing process.4. The method of claim 3 , wherein said batch feeding occurs at least once every day during the culturing process.5. The method of any one of the preceding claims claim 3 , wherein said batch feeding is terminated at least 10 days after initiation of the culturing process.6. The method of any one of the preceding claims claim 3 , wherein said batch feeding comprises at least two separate feeds.7. The method of claim 6 , wherein the ratio of feeding of the first feed to the second feed is about 10:1.8. The method of any one of the preceding claims claim 6 , wherein the addition of supplemental zinc occurs at least once during the culturing process.9. The method of any one of the preceding claims claim 6 , wherein the addition of supplemental zinc occurs about 11 days after initiation of the culturing process.10. The method of any one of the preceding claims claim 6 , wherein said quantity of supplemental zinc is between about 60 to 100 μM zinc.11. The method of claim 2 , wherein said quantity of supplemental zinc is between about 60 to 100 μM ...

METHOD, DEVICE AND KIT FOR MASS CULTIVATION OF CELLS USING POLYIMIDE POROUS MEMBRANE

The present invention pertains to a method for the mass cultivation of cells, and a cell cultivation device and kit. The present invention further pertains to a continuous cell cultivation method and a continuous cell cultivation device in which a carrier is used. 2. The cell culturing apparatus according to claim 1 , wherein the culturing unit further comprises an air supply port and an air discharge port claim 1 , or an oxygen exchange membrane.3. The cell culturing apparatus according to claim 2 , wherein the air supply port and the air discharge port are claim 2 , respectively claim 2 , a 5% COgas-containing air supply port and a 5% COgas-containing air discharge port.4. The cell culturing apparatus according to claim 1 , wherein the culturing unit further houses means for agitating the porous polyimide film.5. The cell culturing apparatus according to claim 1 , wherein the culturing unit further comprises a culture medium discharge line claim 1 , the first end of the culture medium discharge line being connected to the culture medium housing vessel claim 1 , the second end of the culture medium discharge line being in communication with the culturing unit interior via the culture medium discharge port of the culturing unit claim 1 , and the medium being able to circulate through the culture medium-supply unit and the culturing unit.6. The cell culturing apparatus according to claim 1 , wherein the culturing unit further comprises a culture medium discharge line claim 1 , the first end of the culture medium discharge line being connected to a culture medium collecting unit and the second end of the culture medium discharge line being in communication with the culturing unit interior via the culture medium discharge port of the culturing unit claim 1 , and the discharged medium can be collected in the culture medium collecting unit.7. The cell culturing apparatus according to claim 1 , further including means for shaking the culturing unit.8. The cell culturing ...

Method for determining process variables in cell cultivation processes

High throughput cultivation systems are used in pharmaceutical research and development. In this connection, samples are taken and analyzed for important parameters using external analysis. The results of the analysis serve to assess the cultivation process and provide important information about the process. Especially with cultivations carried out in parallel, the manual effort of sample preparation is great and can lead to errors. In order to avoid the need for sampling and thus to minimize the errors, a method is described in the present patent application which makes desired target parameters accessible in the form of soft sensors by means of previously recorded process variables. Herein is described a method for determining process-relevant parameters in CHO processes (Chinese hamster ovary) in high-throughput cultivations, in particular glucose, lactate and the live cell density or the live cell volume.

Tetracycline-inducible expression systems

The invention provides inducible promoter systems and their components incorporating components of a tetracycline operon. By coordinating expression of different transcriptional units in these systems as a result of selection of promoters and/or linking the units into the same DNA molecule, these systems can achieve higher levels of expression of coding segments of interest, increased differential levels of expression between on- and off-states, and/or greater responsiveness to inducing agents than conventional systems.

TARGETED INTEGRATION SITES IN CHINESE HAMSTER OVARY CELL GENOME

Described herein are specific CHO genomic sites for targeted insertion of exogenous genes. The sites are located within a sequence selected from SEQ ID NOs: 1-16. 1. An engineered cell , comprising an exogenous nucleic acid molecule inserted in the genome of the engineered cell , wherein the engineered cell is obtained by a process that includes introducing into a host cell a construct for inserting the exogenous nucleic acid molecule into a target site within an expression-enhancing sequence in the genome of the host cell , the expression-enhancing sequence being at least 80% identical to a sequence selected from SEQ ID NOs: 1-16 or a fragment thereof.2. The engineered cell of claim 1 , wherein the construct is a homology recombination construct that includes the exogenous nucleic acid molecule flanked by a first homology arm and a second homology arm claim 1 , the first homology arm being homologous to a sequence upstream of the target site and the second homology arm being homologous to a sequence downstream of the target site.3. The engineered cell of claim 2 , wherein the expression-enhancing sequence is selected from SEQ ID NOs: 1-16.4. The engineered cell of claim 3 , wherein the expression-enhancing sequence is selected from SEQ ID NOs: 2 claim 3 , 4 claim 3 , 6 claim 3 , 8 claim 3 , 10 claim 3 , 12 claim 3 , 14 claim 3 , and 16.5. The engineered cell of claim 1 , wherein the host cell is a CHO cell.6. The engineered cell of claim 5 , wherein the engineered cell contains the exogenous nucleic acid molecule at the target site.7. The engineered cell of claim 5 , wherein the engineered cell contains the exogenous nucleic acid molecule at an off-target site claim 5 , wherein the engineered cell expresses a higher level of the exogenous nucleic acid molecule as compared to a control cell.8. The engineered cell of claim 6 , wherein the expression-enhancing sequence is SEQ ID NO: 7 or 9.9. The engineered cell of claim 6 , wherein the expression-enhancing sequence ...

Truncated actriib-fc fusion proteins

In certain aspects, the present invention provides compositions and methods for modulating (promoting or inhibiting) growth of a tissue, such as bone, cartilage, muscle, fat, brown fat and/or neuronal tissue and for treating metabolic disorders such as diabetes and obesity, as well as disorders associated with any of the foregoing tissue.

Methods of culturing a mammalian cell

Provided herein are methods of culturing a mammalian cell in a liquid medium including poloxamer-188 at a concentration of 1.8 g/L or at a greater concentration than 1.8 g/L more or a liquid medium that includes a poloxamer-188 concentration that is selected based on one or more factors selected from the group of: pore size, pore type, gas flow rate, viable cell density in the medium, and markers related to cell stress.

METHODS OF DECREASING TRISULFIDE BONDS DURING RECOMBINANT PRODUCTION OF POLYPEPTIDES

Provided herein are cell culture media and methods culturing host cells expressing polypeptides to reduce the level of trisulfide bonds in polypeptides produced by the host cells. 1. A method for decreasing trisulfide bond levels in a polypeptide comprising: i) between about 2 μM to about 35 μM iron,', 'ii) between about 0.11 μM to about 2μM riboflavin (vitamin B2),', 'iii) between about 4.5μM to about 80 μM pyridoxine or pyridoxal (vitamin B6),', 'iv) between about 3.4 μM to about 23 μM folic acid (vitamin B9),', 'v) between about 0.2 μM to about 2.5 μM cyanocobalamin (vitamin B12),', 'vi) between about 9 mM and about 10 mM hypotaurine; and', 'vii) between about 0 and about 1.58 mM methionine;, '(a) contacting a host cell comprising a nucleic acid encoding the polypeptide with a basal medium, wherein the basal medium comprises one or more of the following components(b) culturing the host cell to produce the polypeptide; and(c) harvesting the polypeptide produced by the host cell.2. A method for producing a polypeptide , comprising: i) between about 2 μM to about 35 μM iron,', 'ii) between about 0.11 μM to about 2μM riboflavin (vitamin B2),', 'iii) between about 4.5μM to about 80 μM pyridoxine or pyridoxal (vitamin B6),', 'iv) between about 3.4 μM to about 23 μM folic acid (vitamin B9),', 'v) between about 0.2 μM to about 2.5 μM cyanocobalamin (vitamin B12),', 'vi) between about 9 mM and about 10 mM hypotaurine; and', 'vii) between about 0 and about 1.58 mM methionine;, '(a) contacting a host cell comprising a nucleic acid encoding the polypeptide with a basal medium, wherein the basal medium comprises one or more of the following components(b) culturing the host cell to produce the polypeptide; and(c) harvesting the polypeptide produced by the host cell.3. The method of or , whereby the harvested polypeptide has a trisulfide bond level less than a polypeptide produced under identical conditions , except that the concentration of the one or more components differs ...

Methods of cell culture

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using DMSO, are described.

Method For Creating Endometriotic Cells And Endometriosis Model Animal

Provided is a method of producing a cell having properties of endometriosis (hereinafter referred to as “endometriotic-like cell”) from a human mesenchymal stem cell. Also provided are a method of producing an animal model exhibiting human endometriosis, the method including transplanting the cell to an animal, and an animal model of endometriosis. The method of producing an endometriotic-like cell comprises a step of culturing a human mesenchymal stem cell induced to differentiate by allowing a differentiation inducing agent to act on the cell, through use of a low-carbon-source culture medium free of a differentiation inducing agent, in a culture vessel coated with extracellular matrix. 1. A method of producing an endometriotic-like cell , the method comprising a step of culturing a human mesenchymal stem cell induced to differentiate by allowing a differentiation inducing agent to act on the cell , through use of a low-carbon-source culture medium free of a differentiation induction agent , in a culture vessel coated with extracellular matrix.2. A method of producing an endometriotic-like cell , the method comprising the following steps (1) to (3):(1) culturing a human mesenchymal stem cell with a low-carbon-source culture medium to proliferate the cell;(2) culturing the cell obtained in the step (1) with a low-carbon-source culture medium containing a differentiation inducing agent to induce the cell to differentiate; and(3) further culturing the cell obtained in the step (2) with a low-carbon-source culture medium free of a differentiation inducing agent,the culturing of each of the steps (1) to (3) being performed through use of a culture vessel coated with extracellular matrix.3. A method of producing an endometriotic-like cell according to claim 1 , wherein the low-carbon-source proliferation culture medium contains about 500 mg/l to about 3 claim 1 ,000 mg/l of glucose as a carbon source.4. A method of producing an endometriotic-like cell according to claim ...

Methods for preparing antibodies with a defined glycosylation pattern

The present disclosure relates to a process for preparing antibodies with a defined glycosylation pattern, in particular antibodies with a glycan terminating in an N-acetylglucosamine. The antibodies of the disclosure are suitable for use in a process to conjugate a payload thereto. The disclosure also extends to molecules obtained and obtainable from the process disclosed herein, novel molecules and intermediates, compositions comprising said molecules and uses of the molecules and compositions, particularly in treatment, for example in the treatment of cancer.

SAMPLE PREPARATION DEVICE, CELL ANALYZER, AND FILTER MEMBER

A sample preparation device includes: a filter member including a filter configured to separate cells being an analysis target from other components in a sample; a first receptacle and a second receptacle configured to be connected to each other via the filter; a third receptacle capable of holding the sample therein; a negative pressure section configured to apply a negative pressure into the second receptacle, thereby to move the sample in the third receptacle toward the filter via the first receptacle, and thereby to move components other than the analysis target into the second receptacle via the filter; and a positive pressure section configured to apply a positive pressure from the second receptacle side to the filter to which cells being the analysis target are attached. 1. A sample preparation device comprising:a filter member including a filter configured to separate cells being an analysis target from other components in a sample;a first receptacle and a second receptacle configured to be connected to each other via the filter;a third receptacle capable of holding the sample therein;a communication hole formed in the first receptacle and configured to allow the sample to go into and out of the first receptacle;a flow path configured to allow the third receptacle to be communicated with the communication hole;a negative pressure section configured to apply a negative pressure into the second receptacle, thereby to move the sample in the third receptacle toward the filter via the flow path and the first receptacle, and thereby to move components other than the analysis target into the second receptacle via the filter; anda positive pressure section configured to apply a positive pressure from the second receptacle side to the filter to which cells being the analysis target are attached.2. The sample preparation device according to claim 1 , whereinthe first receptacle and the second receptacle are connected to each other via the filter in a liquid tight ...

3D MICROPHYSIOLOGIC SYSTEM

The present invention relates generally to a three-dimensional cell and tissue culture system for the female reproductive tract. In particular provided herein the system includes individual female reproductive cultures in a dynamic microfluidic setting or integrated using a microfluidic microphysiologic system. In some embodiments, the present invention provides ex-vivo female reproductive tract integration in a three dimensional (3D) microphysiologic system. 131-. (canceled)32. A microphysiologic system comprising:(a) a first 3D cell culture subsystem comprising at least a female reproductive cell type in 3D culture and a culture media for said female reproductive cell type; and(b) a second 3D cell culture subsystem comprising at least a non-reproductive system cell type in 3D culture and a culture media for said non-reproductive system cell type;wherein the first 3D cell culture subsystem and the second 3D cell culture subsystem are in unidirectional fluid communication such that fluid from one of the first or second 3D cell culture subsystem flows downstream to the other of the first or second 3D cell culture subsystem.33. The microphysiologic system of claim 32 , wherein the first 3D cell culture subsystem is selected from the group consisting of ovarian claim 32 , fallopian claim 32 , uterine claim 32 , endocervical claim 32 , and ectocervical subsystems.34. The microphysiologic system of claim 33 , wherein the second 3D cell culture subsystem is selected from the group consisting of liver claim 33 , lung claim 33 , breast claim 33 , skin claim 33 , eye claim 33 , adipose claim 33 , bone claim 33 , and blood vessel subsystems.35. The microphysiologic system of claim 34 , wherein the first 3D cell culture subsystem is an ovarian subsystem.36. The microphysiologic system of claim 34 , wherein the first 3D cell culture subsystem is a fallopian subsystem.37. The microphysiologic system of claim 34 , wherein the first 3D cell culture subsystem is a uterine subsystem ...

IMPROVED EUKARYOTIC CELLS FOR PROTEIN MANUFACTURING AND METHODS OF MAKING THEM

Disclosed are mammalian cells and mammalian cell lines that have a reduced load of remnants of past viral/retroviral infections and methods of producing and using the same. 2. The engineered cell of claim 1 , wherein the ERV elements are from gamma retroviral ERVs claim 1 , including Koala epidemic viral (KoRV) claim 1 , Mouse Mammary Tumor Viral (MMTV) claim 1 , Mouse Leukemia Viral (MLV) ERVs and wherein said one or more alterations are adapted to suppress or eliminate release of one or more claim 1 , preferably more than 60% claim 1 , 70% claim 1 , 80% claim 1 , 90% claim 1 , 95% or 100% of said ERVs.3. The engineered cell of claim 2 , wherein said one or more (ERV) elements are or are from a gag claim 2 , po/and/or env gene claim 2 , preferably from the gag gene claim 2 , encoding claim 2 , a MA (matrix) claim 2 , CA (capsid) claim 2 , NC (nucleocapsid) claim 2 , a further domain encoding proteins such as pp12 or p6 and/or are long terminal repeats (LTRs) of a ERV.4. The engineered cell of claim 3 , wherein said one or more ERV elements encode a Gag (groups antigen) protein claim 3 , a Pol (reverse transcriptase) protein and/or an Env (envelope) protein.5. The engineered cell of claim 1 , wherein said deletions claim 1 , additions or substitutions comprise more than 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , 10 claim 1 , 15 claim 1 , 20 claim 1 , 25 claim 1 , 30 claim 1 , 40 claim 1 , 45 claim 1 , 50 claim 1 , 55 claim 1 , 60 claim 1 , 65 claim 1 , 70 claim 1 , 75 claim 1 , 80 claim 1 , 85 claim 1 , 90 claim 1 , 95 claim 1 , 100 claim 1 , 150 or 200 nucleic acids.6. The engineered cell of claim 1 , wherein said alterations are additions in form of targeted integrations and said one or more (ERV) elements are or are from one or more gag claim 1 , po/and/or env genes claim 1 , preferably gag genes claim 1 , and sequences of the gene were subjected to the targeted integrations of a transgene claim 1 , preferably claim 1 , encoding a marker protein such as GFP ...

METHODS OF CELL CULTURE

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using DMSO, are described. 131-. (canceled)32. A method of producing an adalimumab preparation , comprising:(a) providing a cell genetically engineered to express adalimumab; and(b) culturing the cell in a culture medium comprising DMSO under conditions in which the cell expresses adalimumab, to produce a preparation of adalimumab having a target value of one or more of high mannose and fucosylated glycans, wherein the target value of the high mannose glycans is 0.1% to 20% high mannose glycans and the target value of the fucosylated glycans is 70% to 100% fucosylated glycans.331. The method of claim , further comprising harvesting the preparation of adalimumab.34. The method of claim 33 , further comprising formulating the preparation into a drug product.35. The method of claim 32 , wherein the target value is a predetermined pharmaceutical product specification.36. The method of claim 32 , further comprising evaluating the level of one or more of high mannose glycans and fucosylated glycans in the adalimumab preparation.37. The method of claim 32 , wherein the culture medium comprises 0.5% to 5% DMSO.38. The method of claim 37 , wherein the culture medium comprises 1% to 2% DMSO.39. The method of claim 37 , wherein the culture medium comprises 2% to 3% DMSO.40. The method of claim 37 , wherein the culture medium comprises 3% to 4% DMSO.41. The method of claim 37 , wherein the culture medium comprises 2% to 5% DMSO.42. The method of claim 32 , wherein the target value is 5% to 10% high mannose glycans.43. The method of claim 32 , wherein the target value is 10% to 20% high mannose glycans.44. The method of claim 32 , wherein the culturing step comprises a first stage and a second stage.45. The method of claim 44 , wherein the first stage comprises culturing the cell in the culture medium comprising a first level of DMSO claim 44 , and the second stage ...

PRODUCTION OF N-GLYCOPROTEINS FOR ENZYME ASSISTED GLYCOMODIFICATION

The present invention relates to a cell comprising a gene encoding a polypeptide of interest, wherein the polypeptide of interest is expressed comprising one or more posttranslational modification patterns. These modifications are useful for example in improvement of pharmacokinetic properties, i.e. by attaching PEG chains to proteins. The present invention also relates to methods for producing the antibodies and compositions comprising the antibodies, and their uses. 1. A mammalian cell comprising one or more glycosyltransferase genes that have been partially or fully inactivated , and that has more homogeneous glycosylation capacities , wherein mgat2 has been knocked out (KO) , and with or without KO of mgat4A and/or mgat4B and/or mgat5 , allowing production of N-glycans with monoantennary structure.2. The mammalian cell according to claim 1 , further comprising KO of one or more sialyltransferases claim 1 , allowing production of N-glycans with monoantennary structure and without sialic acid capping.3. The mammalian cell according to claim 1 , further comprising KO of one or more sialyltransferases and a B3gnt2 gene claim 1 , allowing production of N-glycans with monoantennary structure and without sialic acid capping and without poly-LacNAc.4. (canceled)5. The mammalian cell according to claim 1 , wherein the cell is derived from Chinese hamster ovary (CHO) or from human kidney.6. The mammalian cell according to claim 1 , wherein the cell is selected from the group consisting of CHO claim 1 , NS0 claim 1 , SP2/0 claim 1 , YB2/0 claim 1 , CHO-K1 claim 1 , CHO-DXB11 claim 1 , CHO-DG44 claim 1 , CHO-S claim 1 , HEK293 claim 1 , HUVEC claim 1 , HKB claim 1 , PER-C6 claim 1 , and NS0 claim 1 , or derivatives of any of these cells.7. The mammalian cell according to claim 1 , wherein the cell is a CHO cell.8. The mammalian cell according to claim 1 , further encoding an exogenous protein.9. (canceled)10. A method for the production of recombinant glycoproteins claim 1 ...

Method for reducing ammonium and lactate production in cho cells

The present invention relates to modified producer cells for improved production of therapeutic proteins. Specifically, the inventors have found that removing genes involved in amino acid catabolism in Chinese Hamster Ovary (CHO) cells improves the cell growth and viability and likely also the yield of a recombinant therapeutic protein produced by the cells.

USE OF GENOMIC NW_006882077.1 IN CHO CELL FOR STABLY EXPRESSING A PROTEIN

Use of genomic NW_006882077.1 in CHO cell for stably expressing a protein is disclosed. The certain site in CHO cell genome for stably expressing a protein is positioned at a base of No. 691045 in a CHO cell gene NW_006882077.1; a sequence of 5′ NNNNNNNNNNNNNNNNNNNNNGG3′ that can be identified by CRISPR/Cas9 technology and positioned in a base range of No. 690980-691090 around the certain site is a target sequence. Various of protein genes are introduced into a fixed site in CHO cell genome, and expressed stably in the present disclosure. 1. A method for stably expressing a protein at a predetermined site in a Chinese hamster ovary (CHO) cell genome , wherein the predetermined site in the CHO cell genome for stably expressing the protein is positioned at a base of No. 691045 in a CHO cell gene NW_006882077.1; the method comprises:identifying a sequence of 5′ NNNNNNNNNNNNNNNNNNNNNGG3′ in a base range of No. 690980-691090 around the predetermined site by CRISPR/Cas9 technology and positioned as a target sequence.2. The method according to claim 1 , wherein the protein has a molecular weight of less than 160 KDa.3. The method according to claim 1 , wherein the protein is one selected from the group consisting of a polypeptide claim 1 , a functional protein claim 1 , an antibody claim 1 , and a fusion protein.4. The method according to claim 1 , wherein the target sequence includes the bases positioned at No. 691068-691090 around the base of No. 691045 in the CHO cell gene NW_006882077.1 claim 1 , and the target sequence is a nucleotide sequence of SEQ ID NO. 1.5. The method according to claim 1 , wherein the target sequence is a nucleotide sequence of SEQ ID NO:2.6. The method according to claim 1 , wherein the target sequence is a nucleotide sequence of SEQ ID NO:3.7. The method according to claim 1 , wherein the target sequence is a nucleotide sequence of SEQ ID NO:4.8. The method according to claim 1 , wherein the target sequence is a nucleotide sequence of SEQ ID ...

Cell Separation and Collection Membrane, and Culturing Sheet, Culturing Device, and Cell Separation and Collection Method Using Same

Provided are a cell separation and collection membrane capable of suppressing non-specific adsorption of cells while specifically adsorbing target cells; a culturing sheet and a culturing apparatus which use the membrane; and a cell separation and collection method. The cell separation and collection membrane included in the culturing sheet comprises: a support; a polymer site fixed to the support and having a changeable structure in accordance with a temperature; a cell adsorbing site bound to the polymer site, exposed relative to an outer surface of the support, and specifically adsorbing a target cell when the cell adsorbing site is brought into contact with a treatment solution containing the target cells; and a hydrophilic site exposed relative to the outer surface and to be brought into contact with the treatment solution. 111.-. (canceled)12. A cell separation and collection membrane comprising:a support;a polymer site fixed to the support and having a changeable structure in accordance with a temperature;a cell adsorbing site bound to the polymer site, exposed relative to an outer surface of the support, and specifically adsorbing a target cell when the cell adsorbing site is brought into contact with a treatment solution containing the target cells; anda hydrophilic site exposed relative to the outer surface and to be brought into contact with the treatment solution, whereina position of any one of the cell adsorbing sites is higher than a position of any one of the hydrophilic sites with respect to the support.13. The cell separation and collection membrane according to claim 12 , wherein at least a part of the polymer site includes a spiral structure.14. The cell separation and collection membrane according to claim 12 , wherein the hydrophilic site is bound to the polymer site.15. The cell separation and collection membrane according to claim 12 , wherein the polymer site comprises a polyamino acid.16. The cell separation and collection membrane ...

REGENERATION AND REPAIR OF NEURAL TISSUE FOLLOWING INJURY

Methods, pharmaceutical compositions and kits for regenerating or repairing neural tissue, decreasing apoptosis and improving neurological function following injury using human umbilical cord tissue-derived cells are disclosed. 1. A method of treating a patient having a neurological injury comprising administering to the patient isolated umbilical cord tissue-derived cells in an amount effective to treat the neurological injury , wherein the umbilical cord tissue-derived cells are derived from human umbilical cord tissue substantially free of blood , are capable of self-renewal and expansion in culture , have the potential to differentiate into cells of at least a neural phenotype , and do not express CD117.2. The method of claim 1 , wherein the neurological injury is cerebral ischemia claim 1 , reperfusion following acute ischemia claim 1 , perinatal hypoxic-ischemic injury claim 1 , cardiac arrest claim 1 , intracranial hemorrhage claim 1 , intracranial lesions claim 1 , whiplash or shaken infant syndrome.3. The method of claim 1 , wherein the umbilical cord tissue-derived cells are genetically engineered to produce a gene product that promotes treatment of the neurological injury.4. The method of claim 1 , wherein the cells are administered with at least one other cell type.5. The method of claim 4 , wherein the other cell type is an astrocyte claim 4 , oligodendrocyte claim 4 , neuron claim 4 , neural progenitor claim 4 , neural stem cell or other multipotent or pluripotent stem cell.6. The method of claim 1 , wherein the umbilical cord tissue-derived cells are administered at a pre-determined site in the central or peripheral nervous system of the patient.7. The method of claim 1 , wherein the umbilical cord tissue-derived cells are administered by injection or infusion.8. The method of claim 1 , wherein the umbilical cord tissue-derived cells further express CD10 claim 1 , CD13 claim 1 , CD44 claim 1 , CD73 claim 1 , CD90 claim 1 , PDGFr-alpha and HLA-A claim ...

METHODS OF CELL CULTURE

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described. 135-. (canceled)36. A method of producing an adalimumab preparation having a target value of one or more of fucosylated glycans , galactosylated glycans , high mannose glycans , and sialylated glycans , the method comprising:(a) providing a cell genetically engineered to express adalimumab;(b) culturing the cell in a culture medium comprising 0.1 mg/L to 10 mg/L putrescine under conditions in which the cell expresses the adalimumab; and wherein the target value of fucosylated glycans, galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of fucosylated glycans, galactosylated glycans, or sialylated glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine; or', 'wherein the target value of high mannose glycans is a level at least 10% lower than a level of high mannose glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine., '(c) harvesting a preparation of the adalimumab produced by the cell that meets the target value of the one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans,'}37. The method of claim 36 , wherein the target value is a level of one or more of fucosylated glycans claim 36 , galactosylated glycans claim 36 , high mannose glycans claim 36 , and sialylated glycans in a reference preparation of adalimumab.38. The method of claim 36 , further comprising evaluating a level of one or more of fucosylated glycans claim 36 , galactosylated glycans claim 36 , high mannose glycans claim 36 , and sialylated glycans in the adalimumab preparation.39. The method of claim 36 , wherein the target value is one or more of:(a) 70% to 100% fucosylated glycans;(b) 1% to 95% galactosylated glycans;(c) ...

Methods and Compositions for the Clinical Derivation of an Allogenic Cell and Therapeutic Uses

Various cells, stem cells, and stem cell components, including associated methods of generating and using such cells are provided. In one aspect, for example, an isolated cell that is capable of self-renewal and culture expansion and is obtained from a subepithelial layer of a mammalian umbilical cord tissue. Such an isolated cell expresses at least three cell markers selected from CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and does not express at least three cell markers selected from CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR. 1. An isolated cell obtained from a subepithelial layer of a mammalian umbilical cord tissue capable of self-renewal and culture expansion;wherein the isolated cell expresses at least three cell markers selected from the group consisting of CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105; andwherein the isolated cell does not express NANOG and at least five cell markers selected from the group consisting of CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.2. The isolated cell of claim 1 , wherein the isolated cell expresses CD29 claim 1 , CD73 claim 1 , CD90 claim 1 , CD166 claim 1 , SSEA4 claim 1 , CD9 claim 1 , CD44 claim 1 , CD146 claim 1 , and CD105.3. The isolated cell of claim 1 , wherein the isolated cell does not express CD45 claim 1 , CD34 claim 1 , CD14 claim 1 , CD79 claim 1 , CD106 claim 1 , CD86 claim 1 , CD80 claim 1 , CD19 claim 1 , CD117 claim 1 , Stro-1 claim 1 , and HLA-DR.4. The isolated cell of claim 1 , wherein the isolated cell is positive for SOX2.5. The isolated cell of claim 1 , wherein the isolated cell is positive for OCT4.6. The isolated cell of claim 1 , wherein the isolated cell is positive for SOX2 and OCT4.7. The isolated cell of claim 1 , wherein the wherein the isolated cell is capable of differentiation into a cell type selected from the group consisting of adipocytes claim 1 , chondrocytes claim 1 , osteocytes claim 1 , ...

INTEGRATION SITES IN CHO CELLS

The present invention relates to the identification of a genomic integration site for heterologous polynucleotides in Chinese Hamster Ovary (CHO) cells resulting in high RNA and/or protein production. More specifically it relates to CHO cells comprising at least one heterologous polynucleotide stably integrated into the 5100A gene cluster of the CHO genome and to methods for the production of said CHO cells. Further, the invention relates to a method for the production of a protein of interest using said CHO cell and to the use of said CHO cell for producing a protein of interest at high yield. Integration within these specific target regions leads to reliable, stable and high yielding production of an RNA and/or protein of interest, encoded by the heterologous polynucleotide. 1. A Chinese hamster ovary (CHO) cell , comprising at least one heterologous polynucleotide , stably integrated into the S100A gene cluster of the CHO cell genome , whereina) the at least one heterologous polynucleotide is integrated upstream of the S100A3/A4/A5/A6 main gene cluster, into a genomic target region corresponding to the sequence of SEQ ID NO: 1; and/orb) the at least one heterologous polynucleotide is integrated downstream of the S100A3/A4/A5/A6 main gene cluster, into a genomic target region corresponding to the sequence of nucleotides 1 to 15,120 of SEQ ID NO: 2.2. The CHO cell of claim 1 , whereina) the upstream genomic target region corresponds to nucleotides 30 to 19,000 of SEQ ID NO: 1, nucleotides 2,940 to 19,000 of SEQ ID NO: 1, nucleotides 4,740 to 19,000 of SEQ ID NO: 1, nucleotides 6,480 to 19,000 of SEQ ID NO: 1, nucleotides 8,280 to 19,000 of SEQ ID NO: 1, nucleotides 10,020 to 19,000 of SEQ ID NO: 1, or nucleotides 11,820 to 19,000 of SEQ ID NO: 1; and/orb) the downstream genomic target region corresponds to nucleotides 1 to 13,160 of SEQ ID NO: 2, nucleotides 1 to 12,000 of SEQ ID NO: 2 or nucleotides 1 to 10,260 of SEQ ID NO: 2.3. The CHO cell of or claim 1 , ...

Methods of cell culture

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using DMSO, are described.