[특허]Methods for generating stem cell-derived β cells and uses thereof

[미국특허] Methods for generating stem cell-derived β cells and uses thereof 원문보기

IPC분류정보
국가/구분	United States(US) Patent 등록
국제특허분류(IPC7판)	C12N-005/071 A61K-035/39
출원번호	US-0975255 (2015-12-18)
등록번호	US-10190096 (2019-01-29)
발명자 / 주소	Melton, Douglas A. Millman, Jeffrey R.
출원인 / 주소	President and Fellows of Harvard College
대리인 / 주소	Warren, Esq., Lisa M.
인용정보	피인용 횟수 : 0 인용 특허 : 41

초록

Disclosed herein are methods for generating SC-β cells, and isolated populations of SC-β cells for use in various applications, such as cell therapy.

대표청구항 ▼

1. A method for generating stem cell-derived β (SC-β) cells, the method comprising contacting a cell population comprising endocrine progenitor cells under conditions suitable to direct differentiation of said endocrine progenitor cells into said SC-β cells with an effective amount of an agent that decreases the level and/or activity of c-Jun N-terminal kinase (JNK), thereby generating said SC-β cells, wherein the endocrine progenitor cells comprise PDX1+/NKX6.1+/NEUROD1+/insulin+/glucagon−/somatostatin− cells. 2. The method of claim 1, further comprising contacting the endocrine progenitor cells under conditions that promote cell clustering with i) a transforming growth factor β (TGF-β) signaling pathway inhibitor and ii) a thyroid hormone signaling pathway activator to induce in vitro maturation of at least some of the endocrine progenitor cells into SC-β cells. 3. The method of claim 1, wherein the effective amount comprises a concentration between about 0.1 μM and about 110 μM. 4. The method of claim 1, wherein the agent inhibits phosphorylation of JNK. 5. The method of claim 1, wherein the agent is SP600125. 6. The method of claim 1, wherein the agent inhibits JNK interacting protein (JIP). 7. The method of claim 1, wherein the agent inhibits mitogen-activated protein kinase kinase 4 (MKK4). 8. The method of claim 1, wherein the effective amount of the agent comprises 1 μM. 9. The method of claim 1, wherein between at least 5% and 65% of the endocrine progenitor cells in the population differentiate into SC-β cells. 10. The method of claim 1, wherein the level and/or activity of JNK is decreased by at least 70%. 11. The method of claim 1, wherein the level and/or activity of JNK is decreased by at least 90%. 12. The method of claim 1, wherein the level and/or activity of INK is completely inhibited by the agent. 13. The method of claim 1, wherein the agent is SP600125, and wherein an effective amount of SP600125 comprises 1 μM. 14. A method for generating stem cell-derived β (SC-β) cells, the method comprising contacting a cell population comprising endocrine progenitor cells or precursors thereof under conditions suitable to direct differentiation of said endocrine progenitor cells or precursors thereof into said SC-β cells with an effective amount of an agent that decreases level and/or activity of c-Jun N-terminal kinase (JNK), thereby generating said SC-β cells, wherein the agent decreases level and/or activity of mitogen-activated protein kinase kinase 4 (MKK4). 15. A method for generating stem cell-derived β (SC-β) cells, the method comprising contacting a cell population comprising endocrine progenitor cells or precursors thereof under conditions suitable to direct differentiation of said endocrine progenitor cells or precursors thereof into said SC-β cells with an effective amount of an agent that decreases the level and/or activity of c-Jun N-terminal kinase (JNK), thereby generating SC-β cells, wherein between at least 5% and 65% of said endocrine progenitor cells in the population differentiate into said SC-β cells.

이 특허에 인용된 특허 (41) 인용/피인용 타임라인 분석

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D'Amour, Kevin A.; Bang, Anne; Baetge, Emmanuel E., Endocrine precursor cells, pancreatic hormone-expressing cells and methods of production.
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Fisk,Gregory J.; Inokuma,Margaret S., Endoderm cells from human embryonic stem cells.
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Kelly, Olivia; Baetge, Emmanuel E.; Carpenter, Melissa, Expansion of definitive endoderm cells.
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Kelly, Olivia; Baetge, Emmanuel E.; Carpenter, Melissa, Expansion of definitive endoderm cells.
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Penelope E. Roberts ; Jennie Powell Mather, Human pancreatic epithelial progenitor cells and methods of isolation and use thereof.
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Agulnick, Alan D., In vitro differentiation of pluripotent stem cells to pancreatic endoderm cells (PEC) and endocrine cells.
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Fisk,Gregory J.; Inokuma,Margaret S., Islet cells from human embryonic stem cells.
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Zender, Lars; Wuestefeld, Torsten, Medicament for liver regeneration and for treatment of liver failure.
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Mandalam,Ramkumar; Xu,Chunhui, Medium for growing human embryonic stem cells.
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Robins, Allan J; Schulz, Thomas C, Methods and compositions for feeder-free pluripotent stem cell media containing human serum.
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Bodnar, Andrea G.; Chiu, Choy-Pik; Gold, Joseph D.; Inokuma, Margaret; Murai, James T.; West, Michael D., Methods and materials for the growth of primate-derived primordial stem cells in feeder-free culture.
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Terskikh, Alexey; Bajpai, Ruchi, Methods for culture and production of single cell populations of human embryonic stem cells.
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Robins, Allan J.; Schulz, Thomas C., Methods for culturing pluripotent stem cells in suspension using ERBB3 ligands.
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D'Amour, Kevin Allen; Agulnick, Alan D.; Eliazer, Susan; Baetge, Emmanuel E., Methods for identifying factors for differentiating definitive endoderm.
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D'Amour, Kevin Allen; Agulnick, Alan D.; Eliazer, Susan; Baetge, Emmanuel E., Methods for identifying factors for differentiating definitive endoderm.
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Dalton, Stephen; Sheppard, Allan; Jones, Karen; Baetge, E. Edward; D'Amour, Kevin A.; Agulnick, Alan D., Methods for increasing definitive endoderm differentiation of pluripotent human embryonic stem cells with PI-3 kinase inhibitors.
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Agulnick, Alan; D'Amour, Kevin; Baetge, Emmanuel Edward, Methods for increasing definitive endoderm production.
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Agulnick, Alan; D'Amour, Kevin; Baetge, Emmanuel Edward, Methods for increasing definitive endoderm production.
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Kelly, Olivia; Bang, Anne, Methods for purifying endoderm and pancreatic endoderm cells derived from human embryonic stem cells.
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Mitalipova,Maisam; Lyons,Ian, Methods for the culture of human embryonic stem cells on human feeder cells.
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Martinson, Laura; Kroon, Evert; D'Amour, Kevin; Baetge, Emmanuel Edward, Methods of producing pancreatic hormones.
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Martinson, Laura; Kroon, Evert; D'Amour, Kevin; Baetge, Emmanuel Edward, Methods of producing pancreatic hormones.
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Martinson, Laura; Kroon, Evert; D'Amour, Kevin; Baetge, Emmanuel Edward, Methods of producing pancreatic hormones.
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D'Amour, Kevin Allen; Agulnick, Alan D.; Eliazer, Susan; Baetge, Emmanuel E., PDX1 expressing endoderm.
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D'Amour, Kevin Allen; Agulnick, Alan D.; Baetge, Emmanuel E., Preprimitive streak and mesendoderm cells.
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Green, Chad; Yu, Xiaojie; Bang, Anne; Brandon, Eugene; Kelly, Olivia; Agulnick, Alan D.; Baetge, E. Edward; D'Amour, Kevin A.; Schulz, Thomas C.; Robins, Allan J., Stem cell aggregate suspension compositions and methods of differentiation thereof.
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Green, Chad; Yu, Xiaojie; Bang, Anne; Brandon, Eugene; Kelly, Olivia; Agulnick, Alan D; Baetge, E Edward; D'Amour, Kevin A; Schulz, Thomas C; Robins, Allan J, Stem cell aggregate suspension compositions and methods of differentiation thereof.
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Funk, Walter D.; Carpenter, Melissa K.; Gold, Joseph D.; Inokuma, Margaret S.; Xu, Chunhui, cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells.
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[미국특허] Methods for generating stem cell-derived β cells and uses thereof 원문보기