Differentiation of human embryonic stem cells into single hormonal insulin positive cells
AssigneeJanssen Biotech, Inc.
InventorAlireza Rezania
Life of this patent
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Abstract
The present invention provides methods to promote the differentiation of pluripotent stem cells. In particular, the present invention provides methods to produce a population of cells, wherein greater than 10% of the cells in the population express markers characteristic of single hormonal pancreatic beta cells.
Claims25
An in vitro method for the stepwise differentiation of pluripotent cells into a population of cells comprising pancreatic endocrine cells, comprising:
- differentiating the pluripotent cells into foregut endoderm cells;
- and
- differentiating the foregut endoderm cells in a first medium supplemented with a shh inhibitor and a BMP inhibitor, thereby producing the population of cells comprising the pancreatic endocrine cells.
An in vitro method for the stepwise differentiation of pluripotent cells into a population of cells comprising pancreatic endocrine cells, comprising
- differentiating the pluripotent cells into foregut endoderm cells;
- differentiating the foregut endoderm cells in a first medium supplemented with SANT-1 and LDN-193189 to produce foregut precursor cells;
- differentiating the foregut precursor cells in a second medium supplemented with a SANT-1 and LDN-193189 to produce pancreatic endoderm cells and pancreatic endocrine precursor cells;
- differentiating the pancreatic endoderm cells and the pancreatic endocrine precursor cells in a third medium supplemented with SANT-1,
- thereby producing the population of cells comprising pancreatic endocrine cells.
An in vitro method for differentiating human embryonic stem cells into a population of cells comprising pancreatic endocrine cells comprising:
- a) differentiating human embryonic stem cells to produce pancreatic foregut cells;
- b) differentiating the pancreatic foregut cells in a first medium supplemented with a shh inhibitor, a BMP inhibitor, a TGF-B inhibitor, and a retinoid to obtain a population of pancreatic endoderm cells;
- c) differentiating the pancreatic endoderm cells into pancreatic endocrine precursor cells in a second medium supplemented with a shh inhibitor, an ALK inhibitor, and a retinoid;
- and
- d) differentiating the pancreatic endocrine precursor cells into pancreatic endocrine cells in a third medium supplemented with a Shh inhibitor and an ALK inhibitor,
- thereby producing the population of cell comprising the pancreatic endocrine cells.
Dependencies are inferred from the claim text.