EP2038404A2 - Milieu de croissance cellulaire - Google Patents

Milieu de croissance cellulaire

Info

Publication number
EP2038404A2
EP2038404A2 EP07766180A EP07766180A EP2038404A2 EP 2038404 A2 EP2038404 A2 EP 2038404A2 EP 07766180 A EP07766180 A EP 07766180A EP 07766180 A EP07766180 A EP 07766180A EP 2038404 A2 EP2038404 A2 EP 2038404A2
Authority
EP
European Patent Office
Prior art keywords
cell culture
embryonic stem
stem cells
cell
primate embryonic
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP07766180A
Other languages
German (de)
English (en)
Inventor
Miho Dept. Biochemistry and Molecular Biology FURUE
Peter Andrews
Tetsuji Dept Molecular Oral Medicine and Maxilofacial Surgery OKAMOTO
Denry J Mount Desert island marine Laboratory SATO
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Sheffield
Original Assignee
University of Sheffield
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Sheffield filed Critical University of Sheffield
Publication of EP2038404A2 publication Critical patent/EP2038404A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/24—Iron; Fe chelators; Transferrin
    • C12N2500/25—Insulin-transferrin; Insulin-transferrin-selenium
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/38—Vitamins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/44—Thiols, e.g. mercaptoethanol
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/46—Amines, e.g. putrescine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/90—Polysaccharides
    • C12N2501/91—Heparin

Definitions

  • the invention relates to the maintenance of primate embryonic stem cells, preferably human embryonic stem cells (hES), in culture in the absence of feeder cells and serum.
  • hES human embryonic stem cells
  • feeder cells typically fibroblasts which have been treated such that they cannot proliferate (e.g. mitomycin or irradiation treatment).
  • feeder fibroblasts are murine in origin but may be derived from other species
  • stem cell represents a generic group of undifferentiated cells that possess the capacity for self-renewal while retaining varying potentials to form differentiated cells and tissues.
  • Stem cells can be totipotent, pluripotent or multipotent. Derivative stem cells that have lost the ability to differentiate also occur and are termed 'nullipotenf stem cells.
  • a totipotent stem cell is a cell that has the ability to form all the cells and tissues that are found in an intact organism, including the extra-embryonic tissues (i.e. the placenta). Totipotent cells comprise the very early embryo (8 cells) and have the ability to form an intact organism.
  • a pluripotent stem cell is a cell that has the ability to form all tissues found in an intact organism although the pluripotent stem cell cannot form an intact organism.
  • a multipotent cell has a restricted ability to form differentiated cells and tissues.
  • adult stem cells are multipotent stem cells and are the precursor stem cells or lineage restricted stem cells that have the ability to form some cells or tissues and replenish senescing or damaged cells/tissues. Generally they cannot form all tissues found in an organism, although some reports have claimed a greater potential for such 'adult' stem cells than originally thought.
  • Pluripotent embryonic stem cells may be principally derived from two embryonic sources.
  • Cells isolated from the inner cell mass are termed embryonic stem (ES) cells.
  • ES embryonic stem
  • similar cells can be derived from the culture of primordial germ cells isolated from the mesenteries or genital ridges of days 8.5-12.5 post coitum embryos. These are referred to as embryonic germ cells (EG cells).
  • EG cells embryonic germ cells
  • Each of these types of pluripotential cell has a similar developmental potential with respect to differentiation into alternate cell types, but possible differences in behaviour (e.g. with respect to imprinting) have led these cells to be distinguished from one another.
  • the term "pluripotent embryonic stem cell” encompasses both cells derived from the inner cell mass and primordial germ cells.
  • WO96/22362 describes cell lines and growth conditions which allow the continuous proliferation of primate ES cells which exhibit a range of characteristics or markers which are associated with stem cells having pluripotent characteristics.
  • WO96/22362 discloses a method of maintaining primate ES cells in culture in an undifferentiated state in the presence of mouse fibroblast feeder cells and serum.
  • hES human ES
  • tissue engineering The potential utility of embryonic stem cells, particularly human ES (hES) cells, in therapeutic tissue engineering is well documented.
  • the pluripotent nature of these cells enables the selection and differentiation of hES cells into any cell/tissue type.
  • adventitious agents such as prions or viruses may infect the recipient when cells exposed to fetal bovine serum or murine feeder cells are used in therapy. It is therefore essential that cell culture of hES cells is conducted to minimise this risk.
  • feeder free and serum free conditions will help reduce this risk.
  • hES cells that have been differentiated into particular cell type derivatives have utility in the identification gene targets for new drugs and existing drugs since the cells are genotypically identical, stable and of known origin.
  • the use of ES cell lines of distinct genotypes also offers possible routes to drug screening and toxicology in a way pertinent to pharmacogenomics.
  • WOO 1/66697 discloses serum free growth of primate ES cells wherein the serum is replaced with fibroblast growth factor, typically human basic fibroblast growth factor (bFGF 4ng/ml).
  • fibroblast growth factor typically human basic fibroblast growth factor (bFGF 4ng/ml).
  • the cell culture media includes KnockOut SR ta (described in WO98/30679 which is incorporated by reference in its entirety) supplemented with bFGF.
  • the cell culture includes irradiated murine fibroblast feeder cells.
  • WO2006/029198 The development of a serum free and feeder free culture method for the growth of hES cells is disclosed in WO2006/029198. These growth conditions use elevated concentrations of bFGF (40-100ng/ml), supplemental agents that include gamma amino butyric acid, pipecholic acid and lithium and including amino acids, lipids, vitamins and glucose. WO2006/029198 also discloses the use of a cell culture substrate comprising human proteins such as fibronectin, vitronectin and laminin.
  • Furue et al discloses the serum and feeder free growth of mouse embryonic stem cells in the presence of leukaemia inhibitory factor (LIF). This is also described in WO2005/063968.
  • LIF leukaemia inhibitory factor
  • a method to maintain a primate embryonic stem cell in cell culture conditions that are cell feeder and serum free comprising: forming a preparation of primate embryonic stem cells in a cell culture vessel comprising cell culture medium that includes fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof and maintaining the primate embryonic stem cells in an undifferentiated state.
  • a method to maintain primate embryonic stem cells in cell culture conditions that are cell feeder free and serum free comprising the steps: i) forming a preparation of primate embryonic stem cells in a cell culture vessel which is coated with a proteinaceous based cell culture support wherein said cells are cultured in a cell culture medium comprising: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof; and
  • pluripotent embryonic stem cells relates to both cells derived from the inner cell mass and primordial germ cells (EG). The possibility also exists of reprogramming somatic or extraembryonic differentiated cells, or more restricted stem cells back to a pluripotent state resembling that of ES cells derived from early embryos.
  • said cells have a stable karyotype.
  • ascorbic acid is ascorbic acid phosphate.
  • said primate embryonic stem cells are pluripotent human embryonic stem cells.
  • said primate embryonic stem cells retain the property to differentiate into at least the endoderm, mesoderm and ectoderm tissues throughout cell culture.
  • fibroblast growth factor is selected from the group consisting of: bFGF/FGF-2, hereinafter acidic FGF/FGF-1, bFGF, FGF-4, FGF-9, FGF-17 or FGF-18.
  • said fibroblast growth factor is bFGF.
  • bFGF is provided at a concentration of between l-50ng/ml; preferably about 10 ng/ml.
  • fibroblast growth factor is recombinant.
  • ascorbic acid phosphate is provided at a concentration of 10-300 ⁇ g/ml; preferably about lOO ⁇ g/ml.
  • 2-ethanolamine is provided at a concentration of 0.05-2. O ⁇ g/ml; preferably about 0.6 ⁇ g/ml.
  • oleic acid is provided at a concentration of 3-15 ⁇ g/ml; preferably about 9.5 ⁇ g/ml.
  • heparin is provided at a concentration of 10- 500ng/ml; preferably about lOOng/ml; preferably, heparin is heparin sulphate salt.
  • said proteinaceous cell culture support is collagen based.
  • the collagen-based cell culture support comprises type I collagen; preferably recombinant type I collagen.
  • said cell culture support comprises recombinant human proteins selected from the group consisting of: collagen I, collagen IV, fibronectin, laminin and vitronectin.
  • said cell support comprises at least two recombinant proteins selected from the group consisting of: collagen I, collagen IV, fibronectin, laminin and vitronectin.
  • said cell support comprises the recombinant proteins collagen I, collagen FV, fibronectin, laminin and vitronectin.
  • said cell culture support is Matrigel 1 " 1 .
  • said primate embryonic stem cells are passaged after addition of EDTA to the cell culture vessel.
  • said primate embryonic stem cells are passaged after addition of collagenase, preferably collagenase IV.
  • primate embryonic stem cells are passaged after addition of dispase.
  • primate embryonic stem cells are passaged after addition of trypsin/EDTA, preferably recombinant trypsin.
  • said primate embryonic stem cells are cloned.
  • the cell culture media does not include the buffering agent HEPES.
  • a method to differentiate primate embryonic stem cells into at least one cell-type in cell culture conditions that are cell feeder free and serum free comprising the steps: i) forming a preparation of primate embryonic stem cells in a cell culture vessel which is coated with a proteinaceous based cell culture support wherein said cells are cultured in a cell culture medium comprising: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor and heparin; and ii) adding an agent that induces the differentiation of the primate embryonic stem cells into at least one cell-type.
  • the primate embryonic stem cells are human pluripotent embryonic stem cells.
  • the cell-type is a neurone.
  • the cell-type is an epithelial cell.
  • said proteinaceous based cell culture support is laminin.
  • a cell culture comprising: primate embryonic stem cells on a proteinaceous based cell culture support and cell culture media comprising: insulin, transferrin, sodium selenite, ethanolamine, 2- mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof.
  • the primate embryonic stem cells are pluripotent human embryonic stem cells.
  • a cell culture comprising: primate embryonic stem cells on a proteinaceous based cell culture support and cell culture medium comprising: insulin, transferrin, sodium selenite, ethanolamine, 2- mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor and heparin characterised in that the cell culture further comprises at least one agent that induces differentiation of the primate embryonic stem cells into at least one cell-type.
  • the primate embryonic stem cells are pluripotent human embryonic stem cells.
  • a cell culture vessel comprising a cell culture medium that includes: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof.
  • said cell culture vessel further comprises primate embryonic stem cells; preferably pluripotent human embryonic stem cells
  • said vessel is selected from the group consisting of: a petri-dish; cell culture bottle or flask; multiwell plate.
  • "Vessel” is construed as any means suitable to contain a primate embryonic stem cell culture.
  • a cell culture medium container comprising a cell culture medium that includes: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor, heparin and ascorbic acid, or ascorbic acid phosphate, or derivative thereof.
  • a cell culture medium container comprising a cell culture media that includes: insulin, transferrin, sodium selenite, ethanolamine, 2-mercaptoethanol, oleic acid complexed with fatty acid free bovine albumin and further wherein the cell culture medium is supplemented with fibroblast growth factor and heparin.
  • Figure 1 illustrates the effect of bFGF on human embryonic stem cell proliferation
  • Figure 2 illustrates the effect of bFGF and heparin on human embryonic stem cell proliferation and morphology
  • Figure 3 illustrates the expression of human embryonic stem cell markers in cells cultured in feeder free conditions
  • Figure 4 illustrates the growth of human embryonic stem cells in various medium
  • Figure 5 illustrates growth curves of human embryonic stem cell-line HUES in feeder free conditions
  • Figure 6 illustrates growth curves of human embryonic stem cell-line Shef 1 in feeder free conditions
  • Table 1 illustrates a summary of cell culture medium components for culturing human embryonic stem in feeder free conditions.
  • Hesf5 medium is identical to Hesf9 medium without the addition of oleic acid complexed with bovine albumin, ascorbic acid phosphate, bFGF, and heparin sulphate.
  • hESF9 medium ESF basal medium without HEPES supplemented with 9 factors, insulin, transferrin, sodium selenite, 2-mercaptoenthanol, 2-ethanolamine, oleic acid complexed with bovine albumin, ascorbic acid phosphate, bFGF, and heparin sulphate. 3. EDTA solution
  • Type I collagen (Nitta Gelatine, Co., Osaka, Japan)
  • the cells were seeded onto 25cm 2 flask coated with 100 ⁇ g/cm2 type I collagen in hESF9 medium.
  • hESF5 medium ESF basal medium without HEPES supplemented with 5 factors, insulin, transferrin, sodium selenite, 2-mercaptoenthanol and 2-ethanolamine.
  • Undifferentiated ES cells are harvested by EDTA solution. 3. Seed the cells onto laminin-coated dish in hESF5 supplemented with 10ng/ml bFGF and lOOng/ml heparin.
  • hESF5 medium supplemented with FA-BSA ESF basal medium without HEPES supplemented with 5 factors, insulin, transferrin, sodium selenite, 2- mercaptoethanol, 2-ethanolamine, and 0.5 mg/ml fatty acid free bovine albumin
  • Undifferentiated ES cells are harvested by EDTA solution.
  • Table 1 Defined medium for feeder and serum free growth (hESF9).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Developmental Biology & Embryology (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Reproductive Health (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un procédé pour cultiver des cellules souches embryonnaires de primate, dans les conditions sans chargeur ni sérum.
EP07766180A 2006-07-12 2007-07-10 Milieu de croissance cellulaire Withdrawn EP2038404A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB0613756.6A GB0613756D0 (en) 2006-07-12 2006-07-12 Cell culture medium
PCT/GB2007/002584 WO2008007082A2 (fr) 2006-07-12 2007-07-10 Milieu de croissance cellulaire

Publications (1)

Publication Number Publication Date
EP2038404A2 true EP2038404A2 (fr) 2009-03-25

Family

ID=36955441

Family Applications (1)

Application Number Title Priority Date Filing Date
EP07766180A Withdrawn EP2038404A2 (fr) 2006-07-12 2007-07-10 Milieu de croissance cellulaire

Country Status (7)

Country Link
EP (1) EP2038404A2 (fr)
JP (1) JP5227318B2 (fr)
CN (1) CN101490243A (fr)
AU (1) AU2007274065A1 (fr)
CA (1) CA2657539A1 (fr)
GB (2) GB0613756D0 (fr)
WO (1) WO2008007082A2 (fr)

Families Citing this family (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2006286149B2 (en) 2005-08-29 2012-09-13 Technion Research And Development Foundation Ltd. Media for culturing stem cells
DK3441459T3 (da) 2006-08-02 2021-06-07 Technion Res & Dev Foundation Fremgangsmåder til ekspansion af embryonale stamceller i en suspensionskultur
US9688956B2 (en) 2008-06-05 2017-06-27 National Cheng Kung University Method for preserving proliferation and differentiation potential of mesenchymal stem cells
CN102076280B (zh) 2008-06-24 2014-08-27 生物活性外科公司 掺入有干细胞或其它生物活性物质的外科缝合线
WO2010017459A1 (fr) 2008-08-07 2010-02-11 Bioactive Surgical, Inc> Revêtements de capture et d’immobilisation de cellules souches pour dispositifs et implants médicaux
CN102317442B (zh) * 2008-12-17 2014-08-13 斯克里普斯研究所 干细胞的产生和保持
US20120190100A1 (en) * 2009-07-21 2012-07-26 Transgene AS Enzymatic composition for the digestion of chicken embryos
DK3633025T3 (da) * 2009-11-12 2022-12-12 Technion Res & Dev Foundation Dyrkningsmedier, cellekulturer og metoder til dyrkning af pluripotente stamceller i en udifferentieret tilstand
CN101812480B (zh) * 2009-11-16 2013-04-10 西北农林科技大学 利用转录因子转染牛体细胞成为诱导性多能干细胞的方法
JP5714267B2 (ja) * 2010-08-26 2015-05-07 日本メナード化粧品株式会社 幹細胞の未分化維持剤及び増殖促進剤
JP6148429B2 (ja) 2011-01-31 2017-06-14 協和発酵バイオ株式会社 ヒト多能性幹細胞の培養方法
CN102586176B (zh) * 2012-01-11 2013-05-08 中国科学院生物物理研究所 一种新型的无动物源、无饲养层的人多能干细胞培养系统
GB201211873D0 (en) * 2012-07-04 2012-08-15 Univ Edinburgh Cell culture
CN104955939B (zh) * 2013-01-31 2018-07-27 味之素株式会社 用于进行多能性干细胞的稳定的未分化维持增殖的培养方法
KR102229266B1 (ko) 2013-05-30 2021-03-19 아지노모토 가부시키가이샤 줄기 세포의 배양용 배지
CN105358707B (zh) * 2013-06-28 2019-06-14 株式会社钟化 筛选多能干细胞生长促进因子的方法
US20160230143A1 (en) * 2013-09-19 2016-08-11 The U.S.A., As Represented By The Secretary, Department Of Health & Human Services Chemically defined culture medium for stem cell maintenance and differentiation
CN104357379B (zh) * 2014-09-30 2017-08-08 刘兴宇 干细胞培养基
CN106032527B (zh) * 2015-03-17 2019-08-13 广州市搏克肿瘤研究所 一种耐受低密度的无饲养层人多能干细胞培养基
CN105907705B (zh) * 2016-06-28 2020-01-24 广州市搏克生物技术有限公司 一种多能干细胞培养基
WO2018132926A1 (fr) * 2017-01-23 2018-07-26 Stemcell Technologies Canada Inc. Milieux et procédés pour améliorer la survie et la prolifération de cellules souches
CN106754715A (zh) * 2017-02-13 2017-05-31 四川新生命干细胞科技股份有限公司 一种用于造血干细胞培养的滋养层的制备方法
CN106754652B (zh) * 2017-03-06 2019-04-02 广州润虹医药科技股份有限公司 iPS细胞分化成外胚层祖细胞的无血清诱导培养基及诱导方法
CN106754657B (zh) * 2017-03-28 2022-07-22 北京赛斯达生物技术有限公司 一种猴胚胎干细胞的无血清培养基
PL3436568T3 (pl) * 2017-05-31 2023-11-20 Promocell Gmbh Podłoże hodowlane do pluripotencjalnych komórek macierzystych
CN110157662A (zh) * 2018-02-15 2019-08-23 郭永珍 一种细胞添加剂及其制备方法
WO2019177420A1 (fr) * 2018-03-16 2019-09-19 사회복지법인 삼성생명공익재단 Composition de milieu comprenant du fgf -17 en tant qu'ingrédient efficace pour favoriser la prolifération de cellules souches et procédé de promotion de la prolifération de cellules souches à l'aide de celle-ci
CN112313329A (zh) * 2018-06-27 2021-02-02 味之素株式会社 干细胞培养用添加物和培养用培养基、以及培养方法
CN110684716A (zh) * 2018-07-08 2020-01-14 郭永珍 一种细胞添加剂的制备方法及其产品
CN109486766B (zh) * 2018-11-26 2021-05-07 中山大学 一种泪腺干细胞、泪腺干细胞的培养体系和培养方法
CN111484970B (zh) * 2020-04-30 2022-09-16 广州再生医学与健康广东省实验室 一种低蛋白含量的无血清无饲养层胚胎与多能干细胞培养基
WO2023150293A2 (fr) * 2022-02-03 2023-08-10 Steakholder Foods Ltd. Formation accélérée de myotube

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998030679A1 (fr) * 1997-01-10 1998-07-16 Life Technologies, Inc. Substitut de serum pour cellules souches embryonnaires
US7439064B2 (en) * 2000-03-09 2008-10-21 Wicell Research Institute, Inc. Cultivation of human embryonic stem cells in the absence of feeder cells or without conditioned medium
JP2005503822A (ja) * 2001-09-28 2005-02-10 イーエス・セル・インターナショナル・プライヴェート・リミテッド 無フィーダーマトリックス上での未分化ヒト胚性幹（ｈｅｓ）細胞の導出および増殖方法ならびにヒトフィーダー層
CA2527847C (fr) * 2003-06-12 2015-09-29 Yeda Research & Development Co. Ltd Amelioration de la differenciation d'oligodendrocytes
ATE466073T1 (de) * 2003-12-26 2010-05-15 Makoto Asashima Basalmedium zur kultivierung von es-zellen
AU2005271723B2 (en) * 2004-07-13 2010-12-16 Asterias Biotherapeutics, Inc. Medium for growing human embryonic stem cells
AU2005282510B2 (en) * 2004-09-08 2010-12-02 Wisconsin Alumni Research Foundation Medium and culture of embryonic stem cells

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2008007082A3 *

Also Published As

Publication number Publication date
AU2007274065A1 (en) 2008-01-17
CN101490243A (zh) 2009-07-22
WO2008007082A2 (fr) 2008-01-17
JP2009542247A (ja) 2009-12-03
CA2657539A1 (fr) 2008-01-17
JP5227318B2 (ja) 2013-07-03
WO2008007082A3 (fr) 2008-03-06
GB0823575D0 (en) 2009-01-28
GB2452456A (en) 2009-03-04
GB0613756D0 (en) 2006-08-23

Similar Documents

Publication Publication Date Title
EP2038404A2 (fr) Milieu de croissance cellulaire
EP3207122B1 (fr) Generation de keratinocytes a partir de cellules souches pluripotentes et entretien de cultures de keratinocytes
JP2020000241A (ja) 多能性幹細胞を培養するための新規な方法および培養培地
US20080241919A1 (en) Defined media for pluripotent stem cell culture
EP2410043A2 (fr) Systemes de culture de cellules souches
WO2008015682A2 (fr) Procédés d'expansion de cellules souches embryonnaires dans une culture en suspension
JP2008201792A (ja) 胚性幹細胞と胚性幹細胞由来の神経前駆細胞
EP1962719A2 (fr) Milieux de culture de cellules souches
CN116194572A (zh) 用于产生间充质干细胞的培养基和方法
IL263557A (en) Differentiation of pluripotent stem cells into iris cells
WO2016187451A1 (fr) Induction multi-voie de la différenciation de cellules souches avec l'arn
EP1841858A2 (fr) Techniques hautement efficaces de survie/proliferation de cellules souches embryonnaires humaines et de survie d'embryons humains en culture
EP1715033A1 (fr) Moyen de preparer des cellules nourricieres pour des cellules souches embryonnaires et des cellules nourricieres
KR102581040B1 (ko) 돼지 만능성 줄기세포 배양용 배지 조성물
JP2023537969A (ja) 胚性幹細胞増殖のための組成物及び方法
CA3221435A1 (fr) Milieux sans serum pour la culture en suspension de cellules souches pluripotentes de betail de mammiferes
KR100856706B1 (ko) 혈관내피 성장인자를 이용한 인간배아 줄기세포로부터도파민 신경세포로의 분화 촉진 방법
OJALA Establishing and optimizing feeder cell-free culture methods for human embryonic stem cells
KR20110130622A (ko) 전분화능 정원줄기세포를 이용한 형질전환 정자의 생산방법

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20081218

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR

AX Request for extension of the european patent

Extension state: AL BA HR MK RS

RIN1 Information on inventor provided before grant (corrected)

Inventor name: SATO, DENRY JMOUNT DESERT ISLAND MARINE LABORATORY

Inventor name: OKAMOTO, TETSULI

Inventor name: ANDREWS, PETER

Inventor name: FURUE, MIHODEPT. BIOCHEMISTRY AND MOLECULAR BIOLOG

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN WITHDRAWN

18W Application withdrawn

Effective date: 20090722