RU2018132199A - Дифференцировка эмбриональных стволовых клеток человека в одногормональные инсулинположительные клетки - Google Patents
Дифференцировка эмбриональных стволовых клеток человека в одногормональные инсулинположительные клетки Download PDFInfo
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- RU2018132199A RU2018132199A RU2018132199A RU2018132199A RU2018132199A RU 2018132199 A RU2018132199 A RU 2018132199A RU 2018132199 A RU2018132199 A RU 2018132199A RU 2018132199 A RU2018132199 A RU 2018132199A RU 2018132199 A RU2018132199 A RU 2018132199A
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- 210000004027 cell Anatomy 0.000 title claims 8
- 230000004069 differentiation Effects 0.000 title 1
- 210000000130 stem cell Anatomy 0.000 title 1
- 238000000034 method Methods 0.000 claims 19
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims 10
- 239000003112 inhibitor Substances 0.000 claims 10
- 235000010323 ascorbic acid Nutrition 0.000 claims 6
- 229960005070 ascorbic acid Drugs 0.000 claims 5
- 239000011668 ascorbic acid Substances 0.000 claims 5
- 210000000496 pancreas Anatomy 0.000 claims 5
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 claims 4
- 210000000936 intestine Anatomy 0.000 claims 4
- 229930002330 retinoic acid Natural products 0.000 claims 4
- 229960001727 tretinoin Drugs 0.000 claims 4
- FOORCIAZMIWALX-ULJHMMPZSA-N (z)-n-(4-benzylpiperazin-1-yl)-1-(3,5-dimethyl-1-phenylpyrazol-4-yl)methanimine Chemical compound CC1=NN(C=2C=CC=CC=2)C(C)=C1\C=N/N(CC1)CCN1CC1=CC=CC=C1 FOORCIAZMIWALX-ULJHMMPZSA-N 0.000 claims 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims 3
- 239000008103 glucose Substances 0.000 claims 3
- CDOVNWNANFFLFJ-UHFFFAOYSA-N 4-[6-[4-(1-piperazinyl)phenyl]-3-pyrazolo[1,5-a]pyrimidinyl]quinoline Chemical group C1CNCCN1C1=CC=C(C2=CN3N=CC(=C3N=C2)C=2C3=CC=CC=C3N=CC=2)C=C1 CDOVNWNANFFLFJ-UHFFFAOYSA-N 0.000 claims 2
- 239000012190 activator Substances 0.000 claims 2
- 210000004039 endoderm cell Anatomy 0.000 claims 2
- RYVZYACBVYKUHD-UHFFFAOYSA-N Alk5 Natural products CC#CC#CCCCCC=CC(=O)NCC(C)C RYVZYACBVYKUHD-UHFFFAOYSA-N 0.000 claims 1
- 102000001893 Bone Morphogenetic Protein Receptors Human genes 0.000 claims 1
- 108010040422 Bone Morphogenetic Protein Receptors Proteins 0.000 claims 1
- 102100028096 Homeobox protein Nkx-6.2 Human genes 0.000 claims 1
- 101000578254 Homo sapiens Homeobox protein Nkx-6.1 Proteins 0.000 claims 1
- 101000578258 Homo sapiens Homeobox protein Nkx-6.2 Proteins 0.000 claims 1
- 101000687905 Homo sapiens Transcription factor SOX-2 Proteins 0.000 claims 1
- 150000000996 L-ascorbic acids Chemical class 0.000 claims 1
- 102100041030 Pancreas/duodenum homeobox protein 1 Human genes 0.000 claims 1
- 101710144033 Pancreas/duodenum homeobox protein 1 Proteins 0.000 claims 1
- 102100024270 Transcription factor SOX-2 Human genes 0.000 claims 1
- 210000003890 endocrine cell Anatomy 0.000 claims 1
- 210000001900 endoderm Anatomy 0.000 claims 1
- 230000009996 pancreatic endocrine effect Effects 0.000 claims 1
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0676—Pancreatic cells
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
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Claims (18)
1. Способ получения одногормональных клеток, включающий:
(a) дифференцировку клеток поджелудочной железы передней кишки в клетки энтодермы поджелудочной железы путем обработки клеток поджелудочной железы передней кишки ингибитором shh, низкой дозой ретиноевой кислоты и аскорбиновой кислотой; и
(b) дифференцировку клеток энтодермы поджелудочной железы в одногормональные эндокринные клетки поджелудочной железы в среде, дополненной ингибитором shh, низкой дозой ретиноевой кислоты и аскорбиновой кислотой.
2. Способ по п.1, в котором клетки энтодермы поджелудочной железы экспрессируют PDX-1, более высокий уровень NKX6.1 и более низкий уровень SOX2 по сравнению с клетками поджелудочной железы передней кишки.
3. Способ по п.1, в котором стадия (а) дополнительно включает обработку активатором PKC и ингибитором рецептора BMP.
4. Способ по п.3, в котором активатор PKC представляет собой TPB.
5. Способ по п. 3 или 4, в котором ингибитор BMP представляет собой LDN-193189.
6. Способ по п.3, в котором стадия (а) включает обработку с использованием 0,25 мкМ SANT-1, 200 нМ TPB, 50 нМ ретиноевой кислоты, 200 нМ TPB, 50 нМ LDN-193189 и 0,1 мМ аскорбиновой кислоты.
7. Способ по п.1, в котором стадия (b) дополнительно включает обработку ингибитором ALK5.
8. Способ по п.7, в котором ингибитор ALK5 представляет собой ингибитор ALK5 II.
9. Способ по п.7, в котором ингибитор ALK5 представляет собой SD208.
10. Способ по п.7, в котором стадия (b) включает обработку с использованием 0,1 мМ аскорбиновой кислоты, 50 нМ ретиноевой кислоты, 0,25 мкМ SANT-1 и 1 мкМ ингибитора Alk5.
11. Способ по любому из пп. 1, 3 или 7, где ингибитор shh на стадии (a), (b) или (a) и (b) представляет собой SANT-1.
12. Способ по любому из пп. 1, 3 или 7, где способ на стадии (a), (b) или (a) и (b) включает обработку с использованием от 0,01 мМ до 1 мМ аскорбиновой кислоты.
13. Способ по любому из пп.1, 3 или 7, где способ на стадии (a), (b) или (a) и (b) включает обработку с использованием от 0,1 мМ до 0,5 мМ аскорбиновой кислоты.
14. Способ по любому из пп.1, 3 или 7, где способ на стадии (a), (b) или (a) и (b) включает обработку в среде, содержащей менее 25 мМ глюкозы.
15. Способ по п.14, в котором концентрация глюкозы находится в диапазоне от 8 мМ до 20 мМ глюкозы.
16. Способ по п.1, в котором клетки поджелудочной железы передней кишки представляют собой клетки поджелудочной железы передней кишки человека.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201161579351P | 2011-12-22 | 2011-12-22 | |
| US61/579,351 | 2011-12-22 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2014130042A Division RU2668798C2 (ru) | 2011-12-22 | 2012-12-07 | Способы in vitro пошаговой дифференцировки полюрипотентных клеток |
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| Publication Number | Publication Date |
|---|---|
| RU2018132199A true RU2018132199A (ru) | 2018-10-31 |
| RU2018132199A3 RU2018132199A3 (ru) | 2019-05-24 |
| RU2705001C2 RU2705001C2 (ru) | 2019-11-01 |
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| Application Number | Title | Priority Date | Filing Date |
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| RU2014130042A RU2668798C2 (ru) | 2011-12-22 | 2012-12-07 | Способы in vitro пошаговой дифференцировки полюрипотентных клеток |
| RU2018132199A RU2705001C2 (ru) | 2011-12-22 | 2012-12-07 | Дифференцировка эмбриональных стволовых клеток человека в одногормональные инсулинположительные клетки |
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| RU2014130042A RU2668798C2 (ru) | 2011-12-22 | 2012-12-07 | Способы in vitro пошаговой дифференцировки полюрипотентных клеток |
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| US (5) | US9388386B2 (ru) |
| EP (1) | EP2794857A4 (ru) |
| JP (5) | JP6441080B2 (ru) |
| KR (2) | KR102203056B1 (ru) |
| CN (1) | CN105143446B (ru) |
| AR (1) | AR089426A1 (ru) |
| AU (3) | AU2012355698B2 (ru) |
| BR (1) | BR112014015417A8 (ru) |
| CA (1) | CA2860107C (ru) |
| HK (1) | HK1203552A1 (ru) |
| MX (1) | MX2014007744A (ru) |
| PH (1) | PH12014501429A1 (ru) |
| RU (2) | RU2668798C2 (ru) |
| SG (2) | SG11201403473QA (ru) |
| WO (1) | WO2013095953A1 (ru) |
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