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RU2018139710A - Дифференцировка плюрипотентных стволовых клеток в клетки кишечной энтодермы средней кишки - Google Patents

Дифференцировка плюрипотентных стволовых клеток в клетки кишечной энтодермы средней кишки Download PDF

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RU2018139710A
RU2018139710A RU2018139710A RU2018139710A RU2018139710A RU 2018139710 A RU2018139710 A RU 2018139710A RU 2018139710 A RU2018139710 A RU 2018139710A RU 2018139710 A RU2018139710 A RU 2018139710A RU 2018139710 A RU2018139710 A RU 2018139710A
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cells
intestinal
intestine
endoderm
intestinal endoderm
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RU2018139710A
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RU2018139710A3 (ru
RU2741114C2 (ru
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Себастьян РИК
Алиреза РЕЗАНИА
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Янссен Байотек, Инк.
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Claims (25)

1. Способ получения популяции клеток кишечной энтодермы средней кишки, включающий культивирование человеческих плюрипотентных стволовых клеток в культуральных средах для индукции дифференцировки в клетки кишечной энтодермы средней кишки, причем получают по существу популяцию клеток кишечной энтодермы средней кишки.
2. Способ получения популяции клеток кишечной энтодермы средней кишки, включающий культивирование человеческих плюрипотентных стволовых клеток в культуральных средах для индукции дифференцировки в клетки кишечной энтодермы средней кишки, причем дифференцированные клетки представляют собой клетки кишечной энтодермы средней кишки, и при этом клетки кишечной энтодермы средней кишки формируются и сохраняют стабильность в виде монослоя.
3. Способ по п. 1 или 2, который включает этапы:
a. культивирования человеческих плюрипотентных стволовых клеток в первой культуральной среде, содержащей GDF-8 и соединение ингибитора GSK3β, в клетки дефинитивной энтодермы;
b. культивирования клеток дефинитивной энтодермы во второй культуральной среде, содержащей аскорбиновую кислоту и FGF7, в клетки примитивной кишечной трубки; и
c. культивирования клеток примитивной кишечной трубки в третьей культуральной среде, содержащей ретиноевую кислоту и BMP2 или BMP4, в клетки кишечной энтодермы средней кишки.
4. Способ по п. 1 или 2, в котором клетки кишечной энтодермы средней кишки экспрессируют CDX2 и FOXA2.
5. Способ по п. 1 или 2, в котором клетки кишечной энтодермы средней кишки экспрессируют транскрипционные факторы, выбранные из группы, состоящей из SOX9, PDX1, KLF5 и HOXC5.
6. Способ по п. 1 или 2, в котором клетки кишечной энтодермы средней кишки не экспрессируют транскрипционные факторы, выбранные из группы, состоящей из SOX2, ALB, PTF1A, HOXA13 и LGR5.
7. Способ по п. 1, в котором клетки кишечной энтодермы средней кишки формируют и поддерживают монослой в культуре.
8. Способ по п. 1 или 2, в котором популяция клеток не содержит мезенхимальных клеток.
9. Способ по п. 1 или 2, в котором популяция клеток не экспрессирует HAND1.
10. Способ по п. 1 или 2, в котором дифференцировку индуцируют in vitro.
11. Способ лечения пациента, страдающего диабетом или подверженного риску его развития, включающий дифференцировку человеческих плюрипотентных стволовых клеток в клетки кишечной энтодермы средней кишки и имплантацию дифференцированных клеток пациенту.
12. Способ по п. 11, дополнительно включающий дифференцировку имплантированных клеток кишечной энтодермы средней кишки in vivo.
13. Способ по п. 11, в котором диабет представляет собой диабет 1-го типа или диабет 2-го типа.
14. Способ по п. 12, в котором клетки кишечной энтодермы средней кишки дополнительно дифференцируются in vivo в энтероэндокринные клетки, причем энтероэндокринные секретируют инкретиновые гормоны.
15. Способ по п. 14, в котором инкретиновые гормоны представляют собой GLP1 и GIP.
16. Способ по п. 11, в котором клетки кишечной энтодермы средней кишки экспрессируют CDX2 и FOXA2.
17. Способ по п. 11, в котором клетки кишечной энтодермы средней кишки экспрессируют транскрипционные факторы, выбранные из группы, состоящей из SOX9, PDX1, KLF5 и HOXC5.
18. Способ по п. 11, в котором клетки кишечной энтодермы средней кишки не экспрессируют транскрипционные факторы, выбранные из группы, состоящей из SOX2, ALB, PTF1A, HOXA13 и LGR5.
19. Способ получения клеток кишечной энтодермы средней кишки, включающий индукцию дифференцировки клеток дефинитивной энтодермы в культуре в клетки примитивной кишечной трубки, причем клетки дефинитивной энтодермы культивируют в культуральных средах, содержащих аскорбиновую кислоту и FGF7.
20. Способ по п. 19, в котором клетки примитивной кишечной трубки культивируют в культуральных средах, содержащих ретиноевую кислоту и BMP2 или BMP4, для дифференцировки в клетки кишечной энтодермы средней кишки.
21. Способ по п. 19, в котором среда для культивирования является кислой.
22. Способ по п. 19, в котором клетки кишечной энтодермы средней кишки формируют и поддерживают монослой в культуре.
RU2018139710A 2016-04-14 2017-04-04 Дифференцировка плюрипотентных стволовых клеток в клетки кишечной энтодермы средней кишки RU2741114C2 (ru)

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US201662322636P 2016-04-14 2016-04-14
US62/322,636 2016-04-14
PCT/US2017/025847 WO2017180361A1 (en) 2016-04-14 2017-04-04 Differentiation of pluripotent stem cells to intestinal midgut endoderm cells

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RU2021100063A Division RU2021100063A (ru) 2016-04-14 2017-04-04 Дифференцировка плюрипотентных стволовых клеток в клетки кишечной энтодермы средней кишки

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