In vitro differentiation of pluripotent stem cells to pancreatic endoderm cells (pec) and endocrine cells
AssigneeViaCyte, Inc.
InventorAlan D. Agulnick
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Abstract
A human immature endocrine cell population and methods for making an immature endocrine cell population are provided. Specifically, immature beta cells and methods for production of immature beta cells are described. Immature beta cells co-express INS and NKX6.1 and are uni-potent and thereby develop into mature beta cells when implanted in vivo. The mature beta cells in vivo are capable of producing insulin in response to glucose stimulation.
Claims19
An in vitro unipotent human immature beta cell in suspension culture expressing INS and NKX6.1 and not substantially expressing NGN3.
A method of producing a unipotent human immature beta cell comprising, contacting human definitive endoderm lineage cells in vitro with a TGFβ superfamily member and a Wnt family member thereby producing a unipotent human immature beta cell.
A method for producing mature beta cells in vivo, said method comprising:
- a. contacting human definitive endoderm lineage cells in vitro with a TGFβ superfamily member and Wnt family member thereby generating immature beta cells;
- b. transplanting the immature beta cells of step (a) into a mammalian subject;
- c. allowing the immature beta cells to mature in vivo to produce a population of cells comprising insulin secreting beta cells.
The method of 10, wherein the human definitive endoderm lineage cells are contacted with at least 50 ng/mL of the TGF˜ superfamily growth factor.
The method of 10, wherein the human definitive endoderm lineage cells are contacted with at least 25 ng/mL of the Wnt family member.
Dependencies are inferred from the claim text.