RU2015137400A - Дифференцировка in vitro плюрипотентных стволовых клеток в панкреатические эндодермальные клетки (пэк) и эндокринные клетки - Google Patents
Дифференцировка in vitro плюрипотентных стволовых клеток в панкреатические эндодермальные клетки (пэк) и эндокринные клетки Download PDFInfo
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- 210000004027 cell Anatomy 0.000 title claims 9
- 238000000338 in vitro Methods 0.000 title claims 6
- 230000004069 differentiation Effects 0.000 title 1
- 210000003890 endocrine cell Anatomy 0.000 title 1
- 210000001778 pluripotent stem cell Anatomy 0.000 title 1
- 238000000034 method Methods 0.000 claims 16
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 claims 14
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims 6
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims 6
- 108060006698 EGF receptor Proteins 0.000 claims 5
- 102000001301 EGF receptor Human genes 0.000 claims 5
- 239000003795 chemical substances by application Substances 0.000 claims 5
- 102100028096 Homeobox protein Nkx-6.2 Human genes 0.000 claims 4
- 101000578254 Homo sapiens Homeobox protein Nkx-6.1 Proteins 0.000 claims 4
- 101000578258 Homo sapiens Homeobox protein Nkx-6.2 Proteins 0.000 claims 4
- 101000603702 Homo sapiens Neurogenin-3 Proteins 0.000 claims 4
- 102100023915 Insulin Human genes 0.000 claims 4
- 102100038553 Neurogenin-3 Human genes 0.000 claims 4
- 230000003213 activating effect Effects 0.000 claims 4
- 239000003102 growth factor Substances 0.000 claims 4
- 101000979190 Homo sapiens Transcription factor MafB Proteins 0.000 claims 2
- 102000048238 Neuregulin-1 Human genes 0.000 claims 2
- 108090000556 Neuregulin-1 Proteins 0.000 claims 2
- 102100023234 Transcription factor MafB Human genes 0.000 claims 2
- 108010023082 activin A Proteins 0.000 claims 2
- 238000001727 in vivo Methods 0.000 claims 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims 2
- 230000035800 maturation Effects 0.000 claims 2
- 101000976075 Homo sapiens Insulin Proteins 0.000 claims 1
- 102000004877 Insulin Human genes 0.000 claims 1
- 108090001061 Insulin Proteins 0.000 claims 1
- 229940125396 insulin Drugs 0.000 claims 1
- 230000003248 secreting effect Effects 0.000 claims 1
- 238000002054 transplantation Methods 0.000 claims 1
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- A61K35/37—Digestive system
- A61K35/39—Pancreas; Islets of Langerhans
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Claims (22)
1. Унипотентная человеческая незрелая бета-клетка in vitro, экспрессирующая INS и NKX6.1, и по существу не экспрессирующая NGN3.
2. Унипотентная человеческая незрелая бета-клетка по п. 1, способная к созреванию в зрелую бета-клетку.
3. Унипотентная человеческая незрелая бета-клетка по п. 1, дополнительно экспрессирующая MAFB.
4. Способ получения унипотентной человеческой незрелой бета-клетки, включающий обеспечение контакта клеток человеческой дефинитивной эндодермальной линии in vitro с членом суперсемейства TGFβ и членом семейства Wnt, с получением, таким образом, унипотентной человеческой незрелой бета-клетки.
5. Способ по п. 4, где указанная унипотентная человеческая незрелая бета-клетка экспрессирует INS, NKX6.1 и по существу не экспрессирует NGN3.
6. Способ по п. 4, дополнительно включающий обеспечение контакта клеток человеческой дефинитивной эндодермальной линии in vitro с агентом, активирующим киназу рецептора ERBB.
7. Способ по п. 4, где фактором роста суперсемейства TGFβ является активин А.
8. Способ по п. 4, где членом семейства Wnt является Wnt3a.
9. Способ по п. 6, где агентом, активирующим киназу рецептора ERBB, является херегулин.
10. Способ получения зрелых бета-клеток in vivo, включающий:
(a) обеспечение контакта клеток человеческой дефинитивной эндодермальной линии in vitro с членом суперсемейства TGFβ и членом семейства Wnt, с получением, таким образом, незрелых бета-клеток;
(b) трансплантацию незрелых бета-клеток с этапа (а) млекопитающему субъекту; и
(c) обеспечение созревания незрелых бета-клеток in vivo с получением популяции клеток, включающих инсулин-секретирующие бета-клетки.
11. Способ по п. 10, дополнительно включающий обеспечение контакта клеток человеческой дефинитивной эндодермальной линии in vitro с агентом, активирующим киназу рецептора ERBB.
12. Способ по п. 10, где фактором роста суперсемейства TGFβ является активин А.
13. Способ по п. 10, где членом семейства Wnt является Wnt3a.
14. Способ по п. 11, где агентом, активирующим киназу рецептора ERBB, является херегулин.
15. Способ по п. 10, где клетки человеческой дефинитивной эндодермальной линии контактируют по меньшей мере с 50 нг/мл фактора роста суперсемейства TGFβ.
16. Способ по п. 10, где клетки человеческой дефинитивной эндодермальной линии контактируют по меньшей мере с 25 нг/мл члена семейства Wnt.
17. Способ по п. 11, где клетки человеческой дефинитивной эндодермальной линии контактируют с количеством агента, активирующим киназу рецептора ERBB, в 5-15 раз меньшим, чем количество фактора роста суперсемейства TGFβ и члена семейства Wnt.
18. Способ по п. 10, где незрелые бета-клетки экспрессируют INS и NKX6.1, и по существу не экспрессируют NGN3.
19. Способ по п. 10, где незрелые бета-клетки экспрессируют INS, NKX6.1 и MAFB, и по существу, не экспрессируют NGN3.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
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US201361781005P | 2013-03-14 | 2013-03-14 | |
US61/781,005 | 2013-03-14 | ||
US14/106,330 | 2013-12-13 | ||
US14/106,330 US8859286B2 (en) | 2013-03-14 | 2013-12-13 | In vitro differentiation of pluripotent stem cells to pancreatic endoderm cells (PEC) and endocrine cells |
PCT/US2014/026529 WO2014160413A1 (en) | 2013-03-14 | 2014-03-13 | In vitro differentiation of pluripotent stem cells to pancreatic endoderm cells (pec) and endocrine cells |
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RU2018134644A Division RU2734657C2 (ru) | 2013-03-14 | 2014-03-13 | Дифференцировка in vitro плюрипотентных стволовых клеток в панкреатические эндодермальные клетки (пэк) и эндокринные клетки |
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