Patent ID: 11920160
Assignee: CEDARS-SINAI MEDICAL CENTER
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC A  C

Claim 0:
1. A method of differentiating a human induced pluripotent stem cell (iPSC) into a mature insulin producing cell comprising:
(a) providing a quantity of human iPSCs;
(b) culturing the human iPSCs in the presence of activin A at 100 ng/mL and wingless-related MMTV integration site 3A (WNT3A) at 25 ng/mL for 3 days to generate definitive endoderm cells that are positive for C-X-C motif chemokine receptor 4 (CXCR4) and SRY-box transcription factor 17 (SOX17);
(c) further culturing the definitive endoderm cells with endothelial cells in a collagen I, collagen IV, and laminin I gel mixture, in the presence of all-trans retinoic acid (RA) at 2 μmol/L and keratinocyte growth factor (KGF) at 50 ng/mL for about 6 days;
(d) additionally culturing in the presence of epidermal growth factor (EGF) at 50 ng/mL for 5 or 6 days; and
(e) further culturing in the presence of hepatocyte growth factor (HGF) at 50 ng/mL, insulin-like growth factor (IGF1) at 50 ng/mL, exendin-4 at 50 ng/mL, and nicotinamide at 10 mmol/L for about 7 days, thereby obtaining the mature insulin producing cells that secrete insulin and/or human C-peptide in response to glucose stimulation, and wherein the mature insulin producing cells express urocortin 3, pancreatic duodenal homeobox 1 (PDX-1), and C-peptide.