DIFFERENTIATION OF HUMAN EMBRYONIC STEM CELLS
First Claim
1. A method for generating cells expressing markers characteristic of the pancreatic endocrine lineage, comprising differentiating cells expressing markers characteristic of the pancreatic endoderm lineage into cells expressing markers characteristic of the pancreatic endocrine lineage by treating the cells expressing markers characteristic of the pancreatic endoderm lineage with a gamma secretase inhibitor, or Exendin-4, or Exendin 4 and HGF, in medium supplemented with glucose at a concentration from about 10 mM to about 20 mM.
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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.
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Citations
31 Claims
- 1. A method for generating cells expressing markers characteristic of the pancreatic endocrine lineage, comprising differentiating cells expressing markers characteristic of the pancreatic endoderm lineage into cells expressing markers characteristic of the pancreatic endocrine lineage by treating the cells expressing markers characteristic of the pancreatic endoderm lineage with a gamma secretase inhibitor, or Exendin-4, or Exendin 4 and HGF, in medium supplemented with glucose at a concentration from about 10 mM to about 20 mM.
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16. A method for differentiating pluripotent stem cells into cells expressing markers characteristic of the pancreatic endocrine lineage, comprising the steps of:
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a. Culturing the pluripotent stem cells, b. Differentiating the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage, by treating the pluripotent cells with activin A, c. Differentiating the cells expressing markers characteristic of the definitive endoderm lineage into cells expressing markers characteristic of the pancreatic endoderm lineage, by treating the cells expressing markers characteristic of the definitive endoderm lineage with at least one fibroblast growth factor, or with retinoic acid and at least one fibroblast growth factor, and d. Differentiating the cells expressing markers characteristic of the pancreatic endoderm lineage into cells expressing markers characteristic of the pancreatic endocrine lineage, by treating the cells expressing markers characteristic of the pancreatic endoderm lineage with a gamma secretase inhibitor, or Exendin-4, or Exendin 4 and HGF, in medium supplemented with glucose at a concentration from about 10 mM to about 20 mM. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28, 29, 30, 31)
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Specification