WO2019144606A1 - Procédé de criblage et de différenciation de hpsc en msc - Google Patents

Procédé de criblage et de différenciation de hpsc en msc Download PDF

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Publication number
WO2019144606A1
WO2019144606A1 PCT/CN2018/101081 CN2018101081W WO2019144606A1 WO 2019144606 A1 WO2019144606 A1 WO 2019144606A1 CN 2018101081 W CN2018101081 W CN 2018101081W WO 2019144606 A1 WO2019144606 A1 WO 2019144606A1
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Prior art keywords
culture
cells
mscs
differentiation
gelatin
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PCT/CN2018/101081
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English (en)
Chinese (zh)
Inventor
王淋立
李强
陈月花
关春燕
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皓昇莱生物制药有限公司
王淋立
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Publication of WO2019144606A1 publication Critical patent/WO2019144606A1/fr

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    • 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
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
    • C12N5/0668—Mesenchymal stem cells from other natural sources
    • 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
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
    • 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
    • C12N2501/15—Transforming growth factor beta (TGF-β)
    • 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
    • C12N2501/16—Activin; Inhibin; Mullerian inhibiting substance
    • 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
    • C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
    • C12N2506/02—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from embryonic cells
    • 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
    • C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
    • C12N2506/45—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from artificially induced pluripotent stem cells
    • 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
    • C12N2510/00—Genetically modified cells

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  • Figure 7 shows MSCs cells differentiated from hESCs into phenotypic mature, spindle-shaped forms.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Rheumatology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de criblage et de différenciation de hPSC en MSC, comprenant les étapes suivantes : 1) transfert de souches de hPSC indifférenciées sur une plaque de culture revêtue d'une matrice extracellulaire pour une culture ; 2) digestion de cellules, et transfert des cellules sur une plaque de culture revêtue d'une matrice extracellulaire de gélatine pour le criblage et la culture des cellules ; et 3) digestion de cellules, et transfert des cellules sur une plaque de culture adhérente pour continuer à cultiver les cellules pour obtenir des MSC. Le procédé raccourcit le temps de culture de MSC et facilite la culture à grande échelle de MSC.
PCT/CN2018/101081 2018-01-26 2018-08-17 Procédé de criblage et de différenciation de hpsc en msc WO2019144606A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810075894.9 2018-01-26
CN201810075894.9A CN108570447A (zh) 2018-01-26 2018-01-26 一种筛选分化hPSCs向MSCs的方法

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WO2019144606A1 true WO2019144606A1 (fr) 2019-08-01

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CN (1) CN108570447A (fr)
WO (1) WO2019144606A1 (fr)

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CN113462642A (zh) * 2021-08-12 2021-10-01 呈诺再生医学科技(珠海横琴新区)有限公司 间充质干细胞的快速诱导分化方法、试剂盒及其应用

Citations (1)

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Publication number Priority date Publication date Assignee Title
CN106520687A (zh) * 2016-10-12 2017-03-22 广东艾时代生物科技有限责任公司 一种诱导多能干细胞向间充质干细胞分化的方法

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EP1937801A1 (fr) * 2005-09-02 2008-07-02 Agency for Science, Technology and Research Procédé de dérivation de cellules souches mésenchymateuses
WO2011091475A1 (fr) * 2010-01-28 2011-08-04 Australian Stem Cell Centre Limited Méthode de différenciation de cellules souches
CA2876512C (fr) * 2012-07-11 2022-12-13 Imstem Biotechnology, Inc. Cellules souches de type mesenchymateuses issues de cellules souches embryonnaires humaines, leurs procedes et leurs utilisations
CN105154393A (zh) * 2014-12-22 2015-12-16 浙江大学 一种胚胎干细胞分化间充质干细胞的方法
CN106554936B (zh) * 2015-09-30 2020-05-05 海门雨霖细胞科技有限责任公司 诱导人干细胞向肝细胞定向分化的新方法
CN105754936B (zh) * 2016-02-25 2018-06-08 付清玲 人诱导性多能干细胞诱导为间充质干细胞的方法

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106520687A (zh) * 2016-10-12 2017-03-22 广东艾时代生物科技有限责任公司 一种诱导多能干细胞向间充质干细胞分化的方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TRAN, N.T. ET AL.: "Efficient Differentiation of Human Pluripotent Stem Cells into Mesenchymal Stem Cells by Modulating Intracellular Signaling Pathways in a Feeder/Serum- Free System", STEM CELLS AND DEVELOPMENT, vol. 21, no. 7, 27 July 2011 (2011-07-27), XP055047708, ISSN: 1547-3287, doi:10.1089/scd.2011.0346 *

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