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RU2017125728A - Культивирование плюрипотентных стволовых клеток в суспензии - Google Patents

Культивирование плюрипотентных стволовых клеток в суспензии Download PDF

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RU2017125728A
RU2017125728A RU2017125728A RU2017125728A RU2017125728A RU 2017125728 A RU2017125728 A RU 2017125728A RU 2017125728 A RU2017125728 A RU 2017125728A RU 2017125728 A RU2017125728 A RU 2017125728A RU 2017125728 A RU2017125728 A RU 2017125728A
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cells
expression
endoderm
ptf1a
differentiating
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RU2017125728A
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RU2017125728A3 (ru
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Бенджамин ФРАЙЕР
Даина ЛАНИАУСКАС
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Янссен Байотек, Инк.
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Claims (7)

1. Способ дифференцировки человеческих плюрипотентных клеток, включающий стадии: дифференцировки клеток энтодермы передней кишки в клетки панкреатической энтодермы посредством культивирования клеток энтодермы передней кишки в динамической суспензионной культуре при рН от около 7,2 до около 7,0 в течение, по меньшей мере, около 24 часов.
2. Способ по п. 1, дополнительно включающий культивирование клеток энтодермы передней кишки в культуре, в которой концентрация клеток равна или превышает около 1,5 миллиона клеток/мл.
3. Способ по п. 1, дополнительно включающий культивирование клеток энтодермы передней кишки в культуре, в которой концентрация клеток превышает или равна около 2,0 миллиона клеток/мл.
4. Способ по п. 1, в котором клетки панкреатической энтодермы являются по существу отрицательными в отношении экспрессии PTF1A и NGN3.
5. Способ по п. 4, дополнительно включающий обогащение клеток панкреатической энтодермы, которые являются по существу отрицательными в отношении экспрессии PTF1A и NGN3, до популяции клеток панкреатической энтодермы, имеющей около 96% или более клеток, которые являются положительными в отношении совместной экспрессии PDX1 и NKX6.1 и положительными в отношении экспрессии PTF1A.
6. Способ по п. 4, дополнительно включающий дифференцировку клеток панкреатической энтодермы, которые являются по существу отрицательными в отношении экспрессии PTF1A и NGN3, до панкреатических эндокринных клеток при отсутствии стадии дифференцировки, на которой формируются клетки, положительные в отношении экспрессии PTF1A.
7. Способ дифференцировки человеческих плюрипотентных клеток, включающий стадии: дифференцировки клеток энтодермы передней кишки в клетки панкреатической энтодермы посредством культивирования клеток энтодермы передней кишки в динамической суспензионной культуре при рН от около 7,2 до около 7,0 в течение, по меньшей мере, около 24 часов, при концентрации клеток, равной или превышающей около 1,5 миллиона клеток/мл и концентрации ретиноида от около 0,5 до около 1,0 мкМ, причем культивирование выполняют при отсутствии компонентов для одного или более из ингибирования, блокирования, активации или стимулирования сигнального пути TGF-бета и сигнального пути BMP и ингибитора сигнального пути белка sonic hedgehog.
RU2017125728A 2014-12-19 2015-12-09 Культивирование плюрипотентных стволовых клеток в суспензии RU2017125728A (ru)

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US201462094509P 2014-12-19 2014-12-19
US62/094,509 2014-12-19
PCT/US2015/064713 WO2016100035A1 (en) 2014-12-19 2015-12-09 Suspension culturing of pluripotent stem cells

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BR112017012974A2 (pt) 2018-01-09
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CA2970935A1 (en) 2016-06-23
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US20160215268A1 (en) 2016-07-28
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CN107250349A (zh) 2017-10-13
AU2015363008B2 (en) 2021-12-09
WO2016100035A1 (en) 2016-06-23
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