Nothing Special   »   [go: up one dir, main page]

RU2016121409A - SUSPENDING AND CLUSTERING OF HUMAN PLURIPOTENT STEM CELLS WITH THE PURPOSE OF THEIR DIFFERENTIATION IN PANCREATIC ENDOCRINE CELLS - Google Patents

SUSPENDING AND CLUSTERING OF HUMAN PLURIPOTENT STEM CELLS WITH THE PURPOSE OF THEIR DIFFERENTIATION IN PANCREATIC ENDOCRINE CELLS Download PDF

Info

Publication number
RU2016121409A
RU2016121409A RU2016121409A RU2016121409A RU2016121409A RU 2016121409 A RU2016121409 A RU 2016121409A RU 2016121409 A RU2016121409 A RU 2016121409A RU 2016121409 A RU2016121409 A RU 2016121409A RU 2016121409 A RU2016121409 A RU 2016121409A
Authority
RU
Russia
Prior art keywords
cells
pluripotent
clusters
medium
differentiating
Prior art date
Application number
RU2016121409A
Other languages
Russian (ru)
Other versions
RU2016121409A3 (en
RU2689710C2 (en
Inventor
Бенджамин ФРАЙЕР
Даина ЛАНИАУСКАС
Марсия БЛЭКМУР
Хайюнь ВАН
Костадинка ЛИЛОВА
Шелли НЕЛЬСОН
Элизабет РОСОЧА
Original Assignee
Янссен Байотек, Инк.
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
Priority claimed from US13/998,974 external-priority patent/US10377989B2/en
Application filed by Янссен Байотек, Инк. filed Critical Янссен Байотек, Инк.
Publication of RU2016121409A publication Critical patent/RU2016121409A/en
Publication of RU2016121409A3 publication Critical patent/RU2016121409A3/ru
Application granted granted Critical
Publication of RU2689710C2 publication Critical patent/RU2689710C2/en

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
    • C12N5/0676—Pancreatic cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • 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
    • C12N5/0068—General culture methods using substrates
    • C12N5/0075—General culture methods using substrates using microcarriers
    • 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
    • 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
    • C12N5/0618—Cells of the nervous system
    • 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
    • C12N5/0657—Cardiomyocytes; Heart 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/02—Atmosphere, e.g. low oxygen conditions
    • 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
    • C12N2501/117—Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
    • 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
    • C12N2501/155—Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/19—Growth and differentiation factors [GDF]
    • 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
    • C12N2501/30—Hormones
    • C12N2501/38—Hormones with nuclear receptors
    • C12N2501/385—Hormones with nuclear receptors of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
    • 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
    • C12N2501/40—Regulators of development
    • C12N2501/41—Hedgehog proteins; Cyclopamine (inhibitor)
    • 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
    • C12N2501/40—Regulators of development
    • C12N2501/415—Wnt; Frizzeled
    • 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
    • C12N2501/40—Regulators of development
    • C12N2501/42—Notch; Delta; Jagged; Serrate
    • 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
    • C12N2501/70—Enzymes
    • C12N2501/72—Transferases [EC 2.]
    • C12N2501/727—Kinases (EC 2.7.)
    • 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
    • C12N2501/999—Small molecules not provided for elsewhere
    • 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
    • C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
    • 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
    • C12N2513/00—3D culture
    • 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
    • C12N2531/00—Microcarriers

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Reproductive Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Cardiology (AREA)
  • Rheumatology (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Plural Heterocyclic Compounds (AREA)

Claims (26)

1. Способ выращивания и дифференцировки плюрипотентных клеток в динамически перемешиваемой суспензионной культуральной системе, включающий культивирование плюрипотентных клеток в плоской прикрепленной культуре до агрегированных кластеров клеток и дифференцировку кластеров плюрипотентных клеток в динамической перемешиваемой суспензионной культуральной системе, в котором стадия дифференцировки включает использование ингибитора Cyp26.1. A method of growing and differentiating pluripotent cells in a dynamically agitated suspension culture system, comprising cultivating pluripotent cells in a flat attached culture to aggregated cell clusters and differentiating clusters of pluripotent cells in a dynamically agitated suspension culture system, wherein the differentiation step involves the use of a Cyp26 inhibitor. 2. Способ по п.1, причем указанный способ увеличивает процент клеток в фазе G0/G1 клеточного цикла.2. The method according to claim 1, wherein said method increases the percentage of cells in the G0 / G1 phase of the cell cycle. 3. Способ по п.1, в котором культивирование включает среду, содержащую от приблизительно 0,1% до приблизительно 2% бычьего сывороточного альбумина.3. The method according to claim 1, in which the cultivation includes a medium containing from about 0.1% to about 2% bovine serum albumin. 4. Способ по п.3, в котором среда дополнительно включает в себя ингибитор Rho-киназы.4. The method according to claim 3, in which the medium further includes an Rho kinase inhibitor. 5. Способ по п.1, в котором плюрипотентные клетки являются прикрепленными.5. The method according to claim 1, in which the pluripotent cells are attached. 6. Способ увеличения процента клеток в фазе G0/G1 клеточного цикла, включающий выращивание плюрипотентных клеток в плоской прикрепленной культуре до агрегированных кластеров клеток в среде, включающей в себя от приблизительно 0,1% до приблизительно 2% бычьего сывороточного альбумина, перенос кластеров плюрипотентных стволовых клеток из плоской прикрепленной культуры в динамическую суспензию и культивирование клеток в динамической суспензии в среде с добавлением малой молекулы и, необязательно, представителя семейства TGFβ.6. A method for increasing the percentage of cells in the G0 / G1 phase of the cell cycle, comprising growing pluripotent cells in a flat attached culture to aggregated cell clusters in a medium comprising from about 0.1% to about 2% bovine serum albumin, transferring pluripotent stem clusters cells from a flat attached culture into a dynamic suspension and culturing the cells in a dynamic suspension in a medium with the addition of a small molecule and, optionally, a member of the TGFβ family. 7. Способ по п.6, в котором малая молекула представляет собой MCX, и причем представитель семейства TGFβ представляет собой GDF-8.7. The method according to claim 6, in which the small molecule is an MCX, and wherein the representative of the TGFβ family is GDF-8. 8. Способ по п.6, в котором выращивание включает среду, содержащую ингибитор Rho-киназы.8. The method according to claim 6, in which the cultivation includes a medium containing an Rho kinase inhibitor. 9. Способ дифференцировки плюрипотентных клеток в дефинитивную энтодерму, включающий выращивание плюрипотентных клеток в плоской прикрепленной культуре до агрегированных кластеров клеток в среде, включающей в себя от приблизительно 0,1% до приблизительно 2% бычьего сывороточного альбумина, перенос кластеров плюрипотентных стволовых клеток из плоской прикрепленной культуры в динамическую суспензию и культивирование клеток в динамической суспензии в среде с добавлением MCX и GDF8 или WNT3A и активина A.9. A method for differentiating pluripotent cells into a definitive endoderm, comprising growing pluripotent cells in a flat attached culture to aggregated cell clusters in a medium comprising from about 0.1% to about 2% bovine serum albumin, transferring pluripotent stem cell clusters from a flat attached culture in a dynamic suspension and culturing cells in a dynamic suspension in a medium supplemented with MCX and GDF8 or WNT3A and activin A. 10. Способ по п.9, в котором выращивание включает среду, содержащую ингибитор Rho-киназы.10. The method according to claim 9, in which the cultivation includes a medium containing an Rho kinase inhibitor. 11. Способ выращивания и дифференцировки плюрипотентных стволовых клеток в динамически перемешиваемой суспензионной культуральной системе, включающий выращивание плюрипотентных стволовых клеток в плоской прикрепленной культуре до агрегированных кластеров клеток, перенос кластеров плюрипотентных стволовых клеток из плоской прикрепленной культуры в динамическую суспензию и дифференцировку плюрипотентных клеток в динамически перемешиваемой суспензионной культуральной системе, причем клетки выращивают до агрегированных кластеров клеток в среде, которая включает в себя от приблизительно 0,1% до приблизительно 2% бычьего сывороточного альбумина.11. A method of growing and differentiating pluripotent stem cells in a dynamically stirred suspension culture system, comprising growing pluripotent stem cells in a flat attached culture to aggregated cell clusters, transferring clusters of pluripotent stem cells from a flat attached culture to a dynamic suspension and differentiating pluripotent cells in a dynamically mixed suspension the culture system, and the cells are grown to aggregate clusters cells in a medium that includes from about 0.1% to about 2% bovine serum albumin. 12. Способ по п.11, в котором кластеры плюрипотентных клеток дифференцируют для создания популяции панкреатических клеток-предшественников, популяции нейронных клеток-предшественников или популяции клеток-предшественников кардиомиоцитов.12. The method of claim 11, wherein the pluripotent cell clusters are differentiated to create a population of pancreatic progenitor cells, a population of neural progenitor cells, or a population of cardiomyocyte progenitor cells. 13. Способ по п.11, в котором плюрипотентные стволовые клетки выбираются из следующих: индуцированные плюрипотентные стволовые клетки, клетки, полученные из ткани пуповины человека, партеноты, человеческие эмбриональные стволовые клетки (клетки hES) и клетки, извлеченные из амниотической жидкости.13. The method according to claim 11, in which the pluripotent stem cells are selected from the following: induced pluripotent stem cells, cells derived from human umbilical cord tissue, parthenotes, human embryonic stem cells (hES cells), and cells extracted from amniotic fluid. 14. Трансплантируемый клеточный продукт, полученный из стволовых клеток, содержащий популяцию дифференцированных панкреатических клеток-предшественников, полученных посредством способа по п.11.14. A transplanted cell product derived from stem cells containing a population of differentiated pancreatic progenitor cells obtained by the method of claim 11. 15. Способ выращивания и дифференцировки плюрипотентных клеток в динамически перемешиваемой суспензионной культуральной системе, включающий:15. A method of growing and differentiating pluripotent cells in a dynamically stirred suspension culture system, including: a. выращивание плюрипотентных клеток в плоской прикрепленной культуре до агрегированных кластеров клеток в среде, включающей в себя от приблизительно 0,1% до приблизительно 2% бычьего сывороточного альбумина;a. growing pluripotent cells in a flat attached culture to aggregated clusters of cells in a medium comprising from about 0.1% to about 2% bovine serum albumin; b. перенос кластеров плюрипотентных клеток с плоской прикрепленной культуры в динамическую суспензионную культуру с использованием фермента или хелатирующего агента;b. the transfer of clusters of pluripotent cells from a flat attached culture in a dynamic suspension culture using an enzyme or chelating agent; c. поддержание кластеров клеток в динамически перемешиваемой суспензионной культуральной системе; иc. maintaining cell clusters in a dynamically stirred suspension culture system; and d. дифференцировку кластеров плюрипотентных клеток в динамически перемешиваемой суспензионной культуральной системе для генерирования популяции панкреатических клеток-предшественников, популяции нейронных клеток-предшественников или популяции клеток-предшественников кардиомиоцитов;d. differentiating clusters of pluripotent cells in a dynamically stirred suspension culture system to generate a population of pancreatic progenitor cells, a population of neural progenitor cells, or a population of cardiomyocyte progenitor cells; причем дифференцировочная среда включает в себя кислород в диапазоне от недостаточности кислорода до приблизительно 30% от уровня окружающей среды, уровень липидов в диапазоне от 0,1% до приблизительно 2% или их комбинацию.moreover, the differentiating medium includes oxygen in the range from oxygen deficiency to about 30% of the ambient level, lipid levels in the range from 0.1% to about 2%, or a combination thereof. 16. Способ по п.15, в котором кластеры стволовых клеток экспрессируют CD9, SSEA4, TRA-1-60 и TRA-1-81, и не экспрессируют CXCR4.16. The method of claim 15, wherein the stem cell clusters express CD9, SSEA4, TRA-1-60, and TRA-1-81, and do not express CXCR4. 17. Способ по п.15, в котором среда для выращивания дополнительно включает ингибитор Rho-киназы.17. The method of claim 15, wherein the growth medium further comprises an Rho kinase inhibitor. 18. Способ по п.15, в котором среда для дифференцировки содержит от приблизительно 0,1% до приблизительно 2% бычьего сывороточного альбумина.18. The method according to clause 15, in which the medium for differentiation contains from about 0.1% to about 2% bovine serum albumin. 19. Способ по п.15, в котором плюрипотентные клетки выбирают из группы, состоящей из индуцированных плюрипотентных стволовых клеток, клеток, полученных из ткани пуповины человека, партенотов, человеческих эмбриональных стволовых клеток (клетки hES) и клеток, извлеченных из амниотической жидкости.19. The method of claim 15, wherein the pluripotent cells are selected from the group consisting of induced pluripotent stem cells, cells derived from human umbilical cord tissue, parthenotes, human embryonic stem cells (hES cells), and cells extracted from amniotic fluid. 20. Способ по п.15, в котором по данному способу генерируются панкреатические клетки-предшественники, экспрессирующие фактор транскрипции β-клеток.20. The method according to clause 15, in which the method generates pancreatic progenitor cells expressing the transcription factor of β-cells. 21. Способ по п.15, в котором факторами транскрипции являются PDX1 и/или NKX6.1. 21. The method according to clause 15, in which the transcription factors are PDX1 and / or NKX6.1.
RU2016121409A 2013-11-01 2014-05-21 Suspending and clustering human pluripotent stem cells for their differentiation into pancreatic endocrine cells RU2689710C2 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US201361962158P 2013-11-01 2013-11-01
US61/962,158 2013-11-01
US13/998,974 2013-12-30
US13/998,974 US10377989B2 (en) 2012-12-31 2013-12-30 Methods for suspension cultures of human pluripotent stem cells
PCT/US2014/038993 WO2015065524A2 (en) 2013-11-01 2014-05-21 Suspension and clustering of human pluripotent stem cells for differentiation into pancreatic endocrine cells

Publications (3)

Publication Number Publication Date
RU2016121409A true RU2016121409A (en) 2017-12-04
RU2016121409A3 RU2016121409A3 (en) 2018-03-29
RU2689710C2 RU2689710C2 (en) 2019-05-29

Family

ID=53004904

Family Applications (2)

Application Number Title Priority Date Filing Date
RU2016121409A RU2689710C2 (en) 2013-11-01 2014-05-21 Suspending and clustering human pluripotent stem cells for their differentiation into pancreatic endocrine cells
RU2016121404A RU2016121404A (en) 2013-11-01 2014-06-17 SUSPENDING AND CLUSTERING OF HUMAN PLURIPOTENT STEM CELLS WITH THE PURPOSE OF THEIR DIFFERENTIATION IN PANCREATIC ENDOCRINE CELLS

Family Applications After (1)

Application Number Title Priority Date Filing Date
RU2016121404A RU2016121404A (en) 2013-11-01 2014-06-17 SUSPENDING AND CLUSTERING OF HUMAN PLURIPOTENT STEM CELLS WITH THE PURPOSE OF THEIR DIFFERENTIATION IN PANCREATIC ENDOCRINE CELLS

Country Status (12)

Country Link
EP (2) EP3063268A4 (en)
JP (2) JP2016534731A (en)
KR (4) KR20180128529A (en)
CN (2) CN105793413A (en)
AU (4) AU2014342995C1 (en)
BR (1) BR112016009393A8 (en)
CA (2) CA2928639A1 (en)
MX (2) MX2016005657A (en)
PH (2) PH12016500783A1 (en)
RU (2) RU2689710C2 (en)
SG (2) SG11201603045VA (en)
WO (2) WO2015065524A2 (en)

Families Citing this family (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105793413A (en) * 2013-11-01 2016-07-20 詹森生物科技公司 Suspension and clustering of human pluripotent stem cells for differentiation into pancreatic endocrine cells
KR20190029587A (en) * 2016-06-23 2019-03-20 티톤 바이오테크, 인크. Cells expressing the parathyroid hormone 1 receptor and uses thereof
WO2018090084A1 (en) * 2016-11-16 2018-05-24 Cynata Therapeutics Limited Pluripotent stem cell assay
US10391156B2 (en) 2017-07-12 2019-08-27 Viacyte, Inc. University donor cells and related methods
EP3654996A4 (en) * 2017-07-21 2021-03-24 Semma Therapeutics, Inc. Re-aggregation of stem cell-derived pancreatic beta cells
JP7000896B2 (en) * 2017-08-23 2022-01-19 王子ホールディングス株式会社 Cell sheet forming member, method for manufacturing cell sheet forming member, and method for manufacturing cell sheet
EP3735457A4 (en) * 2018-01-05 2021-08-25 Platelet Biogenesis, Inc. Compositions and methods for producing megakaryocytes
SG11202006731VA (en) * 2018-01-18 2020-08-28 Agency Science Tech & Res Method for differentiation of human pluripotent stem cell lines in suspension culture
KR102115360B1 (en) * 2018-05-30 2020-05-26 주식회사 바이블리오테카 Adult stem cell culture fluid and a producing method thereof
US20200080107A1 (en) 2018-09-07 2020-03-12 Crispr Therapeutics Ag Universal donor cells
US20220370184A1 (en) 2019-05-31 2022-11-24 W. L. Gore & Associates, Inc. A biocompatible membrane composite
CA3139585C (en) 2019-05-31 2024-01-23 W. L. Gore & Associates, Inc. A biocompatible membrane composite
US20220233299A1 (en) 2019-05-31 2022-07-28 W. L. Gore & Associates, Inc. Cell encapsulation devices with controlled oxygen diffusion distances
WO2020243663A1 (en) 2019-05-31 2020-12-03 W. L. Gore & Associates, Inc. A biocompatible membrane composite
WO2021044379A1 (en) 2019-09-05 2021-03-11 Crispr Therapeutics Ag Universal donor cells
US11116798B2 (en) 2019-09-05 2021-09-14 Crispr Therapeutics Ag Universal donor cells
CN114729323B (en) * 2019-11-22 2024-09-17 诺和诺德股份有限公司 Rotationally aggregated neural microsphere and application thereof
JP2024503290A (en) 2020-12-31 2024-01-25 クリスパー セラピューティクス アクチェンゲゼルシャフト universal donor cells
US20240191185A1 (en) * 2021-03-25 2024-06-13 Kaneka Corporation Method for producing pluripotent stem cell population
CN116218766A (en) * 2023-04-21 2023-06-06 北京意胜生物科技有限公司 Method for preparing pancreatic precursor cells, culture medium and application of pancreatic precursor cells

Family Cites Families (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20050044393A (en) * 2001-11-09 2005-05-12 아르테셀 사이언스, 인크. Endocrine pancreas differentiation of adipose tissue-derived stromal cells and uses thereof
ES2704401T3 (en) * 2006-08-02 2019-03-18 Technion Res & Dev Foundation Methods of embryonic stem cell expansion in a suspension culture
JP5067949B2 (en) * 2006-11-09 2012-11-07 独立行政法人国立国際医療研究センター Method for culturing and subculture of primate embryonic stem cells and method for inducing differentiation thereof
WO2008148105A1 (en) * 2007-05-25 2008-12-04 Medistem Laboratories, Inc. Endometrial stem cells and methods of making and using same
EP2173862B1 (en) * 2007-07-01 2017-04-19 Lifescan, Inc. Single pluripotent stem cell culture
GB0800524D0 (en) * 2008-01-14 2008-02-20 Univ Brighton Cell culture system
JP2011514169A (en) * 2008-03-17 2011-05-06 エージェンシー フォー サイエンス，テクノロジー アンド リサーチ Microcarrier for stem cell culture
MX2011000125A (en) * 2008-06-30 2011-02-25 Centocor Ortho Biotech Inc Differentiation of pluripotent stem cells.
US8895300B2 (en) * 2008-11-04 2014-11-25 Viacyte, Inc. Scalable primate pluripotent stem cell aggregate suspension culture and differentiation thereof
US8008075B2 (en) * 2008-11-04 2011-08-30 Viacyte, Inc. Stem cell aggregate suspension compositions and methods of differentiation thereof
CA2770753C (en) * 2009-08-12 2019-01-15 Kyoto University Method for inducing differentiation of pluripotent stem cells into neural precursor cells
US8932857B2 (en) * 2010-06-15 2015-01-13 Kyoto University Method for selecting reduced differentiation resistance human induced pluripotent stem cells
US9085757B2 (en) * 2010-06-17 2015-07-21 Regents Of The University Of Minnesota Production of insulin producing cells
AU2011296382B2 (en) * 2010-08-31 2016-04-14 Janssen Biotech, Inc. Differentiation of human embryonic stem cells
WO2012117333A1 (en) * 2011-02-28 2012-09-07 Stempeutics Research Malaysia Sdn Bhd Isolation and expansion of adult stem cells, their therapeutic composition and uses thereof
US8951798B2 (en) * 2011-10-13 2015-02-10 Wisconsin Alumni Research Foundation Generation of cardiomyocytes from human pluripotent stem cells
CN105143446B (en) * 2011-12-22 2020-11-03 詹森生物科技公司 Differentiation of human embryonic stem cells into single hormone insulin positive cells
US10519422B2 (en) * 2012-02-29 2019-12-31 Riken Method of producing human retinal pigment epithelial cells
EP2938722B1 (en) * 2012-12-31 2021-12-08 Janssen Biotech, Inc. Suspension and clustering of human pluripotent cells for differentiation into pancreatic endocrine cells
CN105793413A (en) * 2013-11-01 2016-07-20 詹森生物科技公司 Suspension and clustering of human pluripotent stem cells for differentiation into pancreatic endocrine cells

Also Published As

Publication number Publication date
CA2928639A1 (en) 2015-05-07
PH12016500782A1 (en) 2016-06-13
RU2016121409A3 (en) 2018-03-29
SG11201603047PA (en) 2016-05-30
KR20180130001A (en) 2018-12-05
JP2017500013A (en) 2017-01-05
PH12016500783A1 (en) 2016-06-13
CA2928741A1 (en) 2015-05-07
AU2014342995C1 (en) 2018-08-09
EP3063268A2 (en) 2016-09-07
RU2016121404A3 (en) 2018-03-29
KR20160079071A (en) 2016-07-05
WO2015065524A2 (en) 2015-05-07
KR20160079072A (en) 2016-07-05
AU2014342995B2 (en) 2018-04-26
AU2014343007A1 (en) 2016-05-05
WO2015065524A3 (en) 2015-08-13
KR20180128529A (en) 2018-12-03
MX2016005658A (en) 2017-01-05
CN105793413A (en) 2016-07-20
MX2016005657A (en) 2017-01-05
AU2014343007C1 (en) 2018-08-09
AU2014342995A1 (en) 2016-05-05
EP3063269A1 (en) 2016-09-07
JP2016534731A (en) 2016-11-10
SG11201603045VA (en) 2016-05-30
RU2016121404A (en) 2017-12-04
CN105683362A (en) 2016-06-15
AU2018208707A1 (en) 2018-08-16
AU2014343007B2 (en) 2018-04-26
EP3063269A4 (en) 2017-07-12
BR112016009393A8 (en) 2020-03-24
RU2689710C2 (en) 2019-05-29
WO2015065537A1 (en) 2015-05-07
EP3063268A4 (en) 2017-11-29
AU2018208717A1 (en) 2018-08-16

Similar Documents

Publication Publication Date Title
RU2016121409A (en) SUSPENDING AND CLUSTERING OF HUMAN PLURIPOTENT STEM CELLS WITH THE PURPOSE OF THEIR DIFFERENTIATION IN PANCREATIC ENDOCRINE CELLS
CN105358680B (en) Method and composition for the culture entoderm progenitor cells that suspend
RU2015131839A (en) SUSPENDING AND CLUSTERING OF HUMAN PLURIPOTENT CELLS FOR THEIR DIFFERENTIATION IN PANCREATIC ENDOCRINE CELLS
Karlsson et al. Human embryonic stem cell-derived mesenchymal progenitors—potential in regenerative medicine
JP2016534731A5 (en)
AU2013230020B2 (en) Defined media for expansion and maintenance of pluripotent stem cells
Abdelalim et al. Advances and challenges in the differentiation of pluripotent stem cells into pancreatic β cells
MX343462B (en) Cell culture medium.
BR112015015770A2 (en) Human embryonic stem cell culture at air-liquid interface for differentiation in endocrine pancreatic cells
WO2008036447A2 (en) Pluripotent stem cell culture
RU2008120001A (en) PARTENOGENETIC ACTIVATION OF HUMAN OOCYTES TO OBTAIN HUMAN EMBRYONAL STEM CELLS
JP6502323B2 (en) Pluripotent stem cell culture medium
Kunova et al. Adaptation to robust monolayer expansion produces human pluripotent stem cells with improved viability
Lee et al. Y-27632, a ROCK inhibitor, delays senescence of putative murine salivary gland stem cells in culture
De Chiara et al. Characterization of human mesenchymal stem cells isolated from the testis
US20140295552A1 (en) Suspension and clustering of human pluripotent stem cells for differentiation into pancreatic endocrine cells
Fu et al. Autologous feeder cells from embryoid body outgrowth support the long-term growth of human embryonic stem cells more effectively than those from direct differentiation
CN101880649B (en) Stem cell culture method
Balbasi et al. Mouse embryonic stem cell culture in serum-containing or 2i conditions
CN104946581A (en) Special culture medium and method for culturing porcine trophoderm stem cells
Ma et al. Human amniotic fluid stem cells support undifferentiated propagation and pluripotency of human embryonic stem cell without b-FGF in a density dependent manner
CN116769695A (en) Culture medium and method for producing human cells and tissues from teratomas, organoids and embryoid bodies
Şişli et al. Feeder-dependent/independent mouse embryonic stem cell culture protocol
Khoo et al. Autogenic feeder free system from differentiated mesenchymal progenitor cells, maintains pluripotency of the MEL-1 human embryonic stem cells
CN104988115A (en) Application of quinoline or isoquinoline small-molecular compound in culture and cryopreservation of multipotential stem cells of human