Differentiation of human embryonic stem cells
First Claim
1. A method for generating human pancreatic endocrine cells, comprising differentiating isolated pancreatic endoderm cells into the pancreatic endocrine cells by culturing the pancreatic endoderm cells in medium supplemented with glucose at a concentration from about 10 mM to about 20 mM and treating with a factor selected from the group consisting of:
- a gamma secretase inhibitor, Exendin-4, and a combination of Exendin-4 and hepatocyte growth factor.
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Abstract
The present invention provides methods to promote the differentiation of pluripotent stem cells. In particular, the present invention provides an improved method for the formation of pancreatic endoderm, pancreatic hormone expressing cells and pancreatic hormone secreting cells. The present invention also provides methods to promote the differentiation of pluripotent stem cells without the use of a feeder cell layer.
164 Citations
38 Claims
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1. A method for generating human pancreatic endocrine cells, comprising differentiating isolated pancreatic endoderm cells into the pancreatic endocrine cells by culturing the pancreatic endoderm cells in medium supplemented with glucose at a concentration from about 10 mM to about 20 mM and treating with a factor selected from the group consisting of:
- a gamma secretase inhibitor, Exendin-4, and a combination of Exendin-4 and hepatocyte growth factor.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 29, 30, 31, 32, 36, 37, 38)
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16. A method for differentiating human pluripotent stem cells into human pancreatic endocrine cells, comprising the steps of:
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a) culturing the human pluripotent stem cells, b) differentiating the human pluripotent stem cells into human definitive endoderm cells, by treating the human pluripotent cells with activin A, c) differentiating the human definitive endoderm cells into human pancreatic endoderm cells, by treating the human definitive endoderm cells with at least one fibroblast growth factor, or with retinoic acid and at least one fibroblast growth factor, and d) differentiating the human pancreatic endoderm cells into human pancreatic endocrine cells, by culturing the human pancreatic endoderm cells in medium supplemented with glucose at a concentration from about 10 mM to about 20 mM and treating with a factor selected from the group consisting of;
a gamma secretase inhibitor, Exendin-4, and a combination of Exendin-4 and hepatocyte growth factor. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 33, 34, 35)
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Specification