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RU2012153676A - Дифференцирование эмбриональных стволовых клеток человека - Google Patents

Дифференцирование эмбриональных стволовых клеток человека Download PDF

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RU2012153676A
RU2012153676A RU2012153676/10A RU2012153676A RU2012153676A RU 2012153676 A RU2012153676 A RU 2012153676A RU 2012153676/10 A RU2012153676/10 A RU 2012153676/10A RU 2012153676 A RU2012153676 A RU 2012153676A RU 2012153676 A RU2012153676 A RU 2012153676A
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Джин СЮЙ
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Янссен Байотек, Инк.
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Abstract

1. Популяция клеток, экспрессирующих маркеры, характерные для линии панкреатической энтодермы, причем более 50% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.2. Популяция клеток по п. 1, в которой более 60% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.3. Популяция клеток по п. 1, в которой более 70% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.4. Популяция клеток по п. 1, в которой более 80% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.5. Популяция клеток по п. 1, в которой более 90% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.6. Способ генерирования популяции клеток, экспрессирующих маркеры, характерные для линии панкреатической энтодермы, причем более 50% клеток в популяции одновременно экспрессируют PDX-1 и NKX6.1, содержащий следующие стадии:a. культивирование популяции плюрипотентных стволовых клеток;b. дифференцирование популяции плюрипотентных стволовых клеток в популяцию клеток, экспрессирующих маркеры, характерные для линии дефинитивной энтодермы;c. дифференцирование популяции клеток, экспрессирующих маркеры, характерные для линии дефинитивной энтодермы, в популяцию клеток, экспрессирующих маркеры, характерные для линии панкреатической энтодермы, в среде, дополненной активатором протеинкиназы С.7. Способ по п. 6, в котором активатор протеинкиназы С выбран из группы, состоящей из (2S, 5S)-(E, E)-8-(5-(4-(трифторметил)фенил)-2,4-пентадиемоиламино)бензолактама, индолактама V, форбол-12-миристат-13-ацетата и форбол-12,13-дибутирата.8. Способ по п. 6, в котором активатор протеинкиназы С представляет собой (2S, 5S)-(E, E)-8-(5-(4-(трифторметил)фенил)-2,4-пентадиемоиламино)бензолактам.9. Способ по п. 8, в которо�

Claims (20)

1. Популяция клеток, экспрессирующих маркеры, характерные для линии панкреатической энтодермы, причем более 50% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.
2. Популяция клеток по п. 1, в которой более 60% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.
3. Популяция клеток по п. 1, в которой более 70% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.
4. Популяция клеток по п. 1, в которой более 80% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.
5. Популяция клеток по п. 1, в которой более 90% клеток в популяции одновременно экспрессируют PDX1 и NKX6.1.
6. Способ генерирования популяции клеток, экспрессирующих маркеры, характерные для линии панкреатической энтодермы, причем более 50% клеток в популяции одновременно экспрессируют PDX-1 и NKX6.1, содержащий следующие стадии:
a. культивирование популяции плюрипотентных стволовых клеток;
b. дифференцирование популяции плюрипотентных стволовых клеток в популяцию клеток, экспрессирующих маркеры, характерные для линии дефинитивной энтодермы;
c. дифференцирование популяции клеток, экспрессирующих маркеры, характерные для линии дефинитивной энтодермы, в популяцию клеток, экспрессирующих маркеры, характерные для линии панкреатической энтодермы, в среде, дополненной активатором протеинкиназы С.
7. Способ по п. 6, в котором активатор протеинкиназы С выбран из группы, состоящей из (2S, 5S)-(E, E)-8-(5-(4-(трифторметил)фенил)-2,4-пентадиемоиламино)бензолактама, индолактама V, форбол-12-миристат-13-ацетата и форбол-12,13-дибутирата.
8. Способ по п. 6, в котором активатор протеинкиназы С представляет собой (2S, 5S)-(E, E)-8-(5-(4-(трифторметил)фенил)-2,4-пентадиемоиламино)бензолактам.
9. Способ по п. 8, в котором (2S, 5S)-(E, E)-8-(5-(4-(трифторметил)фенил)-2,4-пентадиемоиламино)бензолактам используют в концентрации от приблизительно 20 нМ до приблизительно 500 нМ.
10. Способ по п. 6, в котором в среду, дополненную активатором протеинкиназы С, дополнительно добавили по меньшей мере один фактор из группы, состоящей из фактора, способного ингибировать BMP, ингибитора сигнального каскада рецепторов TGFβ и фактора роста фибробластов.
11. Способ по п. 10, в котором фактор, способный ингибировать BMP, представляет собой Ноггин.
12. Способ по п. 11, в котором Ноггин используют в концентрации от приблизительно 50 нг/мл до приблизительно 500 мкг/мл.
13. Способ по п. 11, в котором Ноггин используют в концентрации приблизительно 100 нг/мл.
14. Способ по п. 10, в котором ингибитор сигнального каскада рецептора TGFβ представляет собой ингибитор ALK5.
15. Способ по п. 14, в котором ингибитор ALK5 представляет собой ингибитор ALK5 II.
16. Способ по п. 15, в котором ингибитор ALK5 II используют в концентрации от приблизительно 0,1 мкМ до приблизительно 10 мкМ.
17. Способ по п. 16, в котором ингибитор ALK5 II используют в концентрации приблизительно 1 мкМ.
18. Способ по п. 10, в котором фактор роста фибробластов представляет собой FGF7.
19. Способ по п. 10, в котором фактор роста фибробластов представляет собой FGF10.
20. Способ по п. 10, в котором фактор роста фибробластов можно использовать в концентрации от приблизительно 50 пг/мл до приблизительно 50 мкг/мл.
RU2012153676/10A 2010-05-12 2011-05-11 Дифференцирование эмбриональных стволовых клеток человека RU2587634C2 (ru)

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US33383110P 2010-05-12 2010-05-12
US61/333,831 2010-05-12
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