Differentiation of human embryonic stem cells into single hormonal insulin positive cells
First Claim
1. An in vitro culture comprising a differentiated population of human pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells and a population of human pancreatic foregut cells in a differentiation medium, wherein the differentiation medium differentiates pancreatic foregut cells into the differentiated population of pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells and wherein said differentiation medium comprises one of the following:
- a retinoid, ascorbic acid, and a shh-inhibitor,a retinoid, ascorbic acid and SANT-1,a retinoid, ascorbic acid, a shh-inhibitor and a TGF-β
ligand,a retinoid, ascorbic acid, a shh-inhibitor and ALK5 inhibitor,a retinoid, ascorbic acid, SANT-1 and a TGF-β
ligand,a retinoid, ascorbic acid, a shh-inhibitor, ALK5 inhibitor, and a BMP inhibitor,a retinoid, ascorbic acid, SANT-1, a TGF-β
ligand and a BMP inhibitor;
ora retinoid, ascorbic acid, a shh-inhibitor, a TGF-β
ligand, and LDN-19318,wherein the presence of ascorbic acid in the differentiation medium increases the number of insulin positive cells and decreases the number of glucagon positive cells and somatostatin positive cells compared to differentiation medium without ascorbic acid.
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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.
181 Citations
35 Claims
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1. An in vitro culture comprising a differentiated population of human pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells and a population of human pancreatic foregut cells in a differentiation medium, wherein the differentiation medium differentiates pancreatic foregut cells into the differentiated population of pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells and wherein said differentiation medium comprises one of the following:
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a retinoid, ascorbic acid, and a shh-inhibitor, a retinoid, ascorbic acid and SANT-1, a retinoid, ascorbic acid, a shh-inhibitor and a TGF-β
ligand,a retinoid, ascorbic acid, a shh-inhibitor and ALK5 inhibitor, a retinoid, ascorbic acid, SANT-1 and a TGF-β
ligand,a retinoid, ascorbic acid, a shh-inhibitor, ALK5 inhibitor, and a BMP inhibitor, a retinoid, ascorbic acid, SANT-1, a TGF-β
ligand and a BMP inhibitor;
ora retinoid, ascorbic acid, a shh-inhibitor, a TGF-β
ligand, and LDN-19318,wherein the presence of ascorbic acid in the differentiation medium increases the number of insulin positive cells and decreases the number of glucagon positive cells and somatostatin positive cells compared to differentiation medium without ascorbic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 25, 28, 29, 30, 31, 32, 33, 34, 35)
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12. An in vitro culture comprising human pancreatic foregut cells in a differentiation medium supplemented with one of the following:
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a retinoid, ascorbic acid, and a shh-inhibitor; a retinoid, ascorbic acid and SANT-1; a retinoid, ascorbic acid, a shh-inhibitor, and a TGF-β
ligand;a retinoid, ascorbic acid, a shh-inhibitor and ALK5 inhibitor; a retinoid, ascorbic acid, SANT-1 and a TGF-β
ligand;a retinoid, ascorbic acid, a shh-inhibitor, ALK5 inhibitor, and a BMP inhibitor; a retinoid, ascorbic acid, SANT-1, a TGF-β
ligand, and a BMP inhibitor;
orwherein the differentiation medium differentiates the human pancreatic foregut cells into a differentiated population of human pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells, and wherein the presence of the ascorbic acid in the differentiation medium increases the number of insulin positive cells and decreases the number of glucagon positive cells and somatostatin positive cells compared to differentiation medium without ascorbic acid. - View Dependent Claims (13, 14, 15, 16, 26)
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- 17. An in vitro culture of a differentiated population of human pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells and a population of human pancreatic foregut cells in a differentiation medium supplemented with a retinoid, ascorbic acid and SANT-1, wherein the differentiation medium differentiates pancreatic foregut cells into the differentiated population of pancreatic endoderm cells in which greater than 10% of the cells are single hormonal insulin positive cells, and wherein the presence of the ascorbic acid in the differentiation medium increases the number of insulin positive cells and decreases the number of glucagon positive cells and somatostatin positive cells compared to differentiation medium without ascorbic acid.
Specification