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

Claim 4:
5. 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) inducing formation of embryoid bodies (EBs) derived from the human iPSCs;
(c) culturing the EBs derived from the human iPSCs with human endothelial cells (ECs) in the presence of activin A at 100 ng/mL and wingless-related MMTV integration site 3A (WNT3A) at 25 ng/mL in a collagen I, collagen IV, and laminin I gel mixture, for at least a day and up to 3 days;
(d) further culturing the EB-EC co-culture derived resulting cells from step (c) in the presence of all-trans retinoic acid (RA) at 2 μmol/L and keratinocyte growth factor (KGF) at 50 ng/mL for 6 days;
(e) additionally culturing in the presence of epidermal growth factor (EGF) at 50 ng/mL for 5 or 6 days; and
(f) 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 7 days,
thereby producing mature insulin producing cells; wherein the mature insulin producing cells 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.