RU2020105325A - Культуральная среда стволовых клеток - Google Patents
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Claims (28)
1. Культуральная среда для культивирования популяции взрослых стволовых эпителиальных клеток, где указанная культуральная среда содержит:
i. агонист Lgr5; и
ii. ингибитор р38.
2. Культуральная среда по п.1, где агонист Lgr5 является любым из R-спондинов 1-4.
3. Культуральная среда по п.1 или 2, где ингибитор р38 выбран из группы, состоящей из SB-202190, SB-203580, VX-702, VX-745, PD-169316, RO-4402257 и BIRB-769.
4. Культуральная среда по любому из предыдущих пунктов, где культуральная среда дополнительно содержит ингибитор TGF-бета.
5. Культуральная среда по п.4, где ингибитор TGF-бета связывается с ALK5, ALK4 и/или ALK7 и снижает их активность.
6. Культуральная среда по п.5, где ингибитор, который связывается и снижает активность ALK5, ALK4 или ALK7, выбран из группы, состоящей из A83-01, SB-431542, SB-505124, SB-525334, SD-208, LY-36494 и SJN-2511.
7. Культуральная среда по любому из предыдущих пунктов, которая содержит один или более дополнительных компонентов, выбранных из: ингибитора BMP, агониста Wnt, лиганда рецептора тирозинкиназы, никотинамида, ингибитора Rock, гастрина, RANKL, ингибитора GSK3, активатора сигнального пути простагландина и тестостерона.
8. Композиция, содержащая культуральную среду по любому из пп.1-7 и внеклеточный матрикс или 3D-матрикс, который имитирует внеклеточный матрикс путем взаимодействия с белками клеточной мембраны, такими как интегрины, например, содержащий ламинин внеклеточный матрикс, такой как MatrigelTM (BD Biosciences).
9. Герметически закрытый сосуд, содержащий культуральную среду или композицию по любому из предыдущих пунктов.
10. Применение культуральной среды по любому из пп.1-7 для увеличения в количестве или дифференцировки стволовой клетки, популяции стволовых клеток, фрагмента ткани или органоида.
11. Способ увеличения в количестве единичной эпителиальной стволовой клетки, популяции эпителиальных стволовых клеток или фрагмента ткани, предпочтительно, для получения органоида, который включает: культивирование единичной эпителиальной стволовой клетки или популяции эпителиальных стволовых клеток в культуральной среде по любому из пп.1-7.
12. Способ по п.11, где способ включает приведение эпителиальной стволовой клетки, популяции эпителиальных стволовых клеток или выделенного фрагмента ткани и культуральной среды в контакт с внеклеточным матриксом или 3D-матриксом, который имитирует внеклеточный матрикс путем его взаимодействия с белками клеточной мембраны, такими как интегрины, например, содержащим ламинин внеклеточным матриксом, таким как MatrigelTM (BD Biosciences).
13. Способ по п.11 или 12, который включает:
культивирование эпителиальной стволовой клетки, популяции эпителиальных стволовых клеток или фрагментов ткани в культуральной среде по любому из пп.1-7;
дальнейшее культивирование эпителиальной стволовой клетки, популяции эпителиальных стволовых клеток или фрагменты ткани и обновление среды средой для дифференциации, которая не содержит одного или нескольких, предпочтительно всех, факторов, выбранных из: ингибитора TGF-бета, ингибитора р38, никотинамида или Wnt.
14. Органоид или популяция клеток, получаемая способом по любому из пп.11-13.
15. Композиция, содержащая:
i) один или несколько органоидов или популяций клеток по п.14; и
ii) питательная среда по любому из пп.1-7 и/или внеклеточный матрикс.
16. Применение органоида по п.14 или популяции клеток по п.14 или композиции по п.8 или 15 для скрининга лекарственных средств, определения мишени, обнаружения мишени, токсикологии, токсикологических скринингов или моделей клеток/органов ex vivo, например, для применения в качестве модели заболевания.
17. Органоид по п.14 или популяция клеток по п.14 или композиция по п.8 или 15 для применения в медицине или диагностике, необязательно, где медицина является персонализированной медициной или регенеративной медициной.
18. Способ скрининга на терапевтическое или профилактическое лекарственное средство или косметическое средство, который включает:
культивирование органоида или популяции клеток по п.14, например, в культуральной среде по пп. 1-7;
воздействие на указанный органоид или популяцию клеток одной молекулой-кандидатом или библиотекой таких молекул-кандидатов;
оценку указанного органоида или популяции клеток на любой эффект, например, на любое изменение в клетке, такое как снижение или утрата пролиферации, морфологические изменения и/или гибель клеток; и
идентификацию молекулы-кандидата, вызывающей указанные эффекты, как перспективного лекарственного или косметического средства.
Applications Claiming Priority (12)
Application Number | Priority Date | Filing Date | Title |
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US201161520569P | 2011-06-10 | 2011-06-10 | |
US61/520,569 | 2011-06-10 | ||
US201161571663P | 2011-06-30 | 2011-06-30 | |
GB1111244.8 | 2011-06-30 | ||
US61/571,663 | 2011-06-30 | ||
GBGB1111244.8A GB201111244D0 (en) | 2011-06-30 | 2011-06-30 | Culture media for stem cells |
US201161513461P | 2011-07-29 | 2011-07-29 | |
US13/194,866 | 2011-07-29 | ||
US61/513,461 | 2011-07-29 | ||
US13/194,866 US9752124B2 (en) | 2009-02-03 | 2011-07-29 | Culture medium for epithelial stem cells and organoids comprising the stem cells |
US201261594295P | 2012-02-02 | 2012-02-02 | |
US61/594,295 | 2012-02-02 |
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