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US 11,890,304 B2Granted
Pancreatic endocrine cells and methods thereof
AssigneeJanssen Biotech, Inc.
InventorJean Xu
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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.
Claims14
A method for generating human pancreatic endocrine cells, comprising:
- differentiating human pluripotent stem cells into definitive endoderm cells by culturing the pluripotent cells in a medium supplemented with activin A, then removing the medium supplemented with activin A;
- differentiating the definitive endoderm cells into pancreatic endoderm cells by culturing the definitive endoderm cells in a medium supplemented with retinoic acid and fibroblast growth factor-2 (FGF-2), then removing the medium supplemented with retinoic acid and FGF-2;
- and
- differentiating the pancreatic endoderm cells into pancreatic endocrine cells by culturing the pancreatic endoderm cells in a medium supplemented with 1S-Benzyl-4R41-(1S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarbamoyl]-2R-hydrozy-5-phenylpentyl] carbamic Acid tert-butyl Ester (L-685,458), thereby generating the pancreatic endocrine cells, wherein FGF-2 is used at a concentration of about 20 ng/ml to about 50 ng/ml.
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