Endocrine precursor cells, pancreatic hormone-expressing cells and methods of production
Disclosed herein are cell cultures and enriched cell populations of endocrine precursor cells, immature pancreatic hormone-expressing cells and mature pancreatic hormone-expressing cells. Also disclosed herein are methods of producing such cell cultures and cell populations.
1. An in vitro method of making a population of human pancreatic and duodenal homeobox (PDX1)-positive pancreatic endoderm, comprising:
a) obtaining a population of human definitive endoderm cells; and
b) contacting the population of human definitive endoderm cells from step (a) with a retinoid, thereby making the population of human PDX1-positive pancreatic endoderm.
2. The method of claim 1 , further comprising contacting the population of human definitive endoderm cells from step (a) with the gamma secretase inhibitor.
3. The method of claim 2 , wherein the gamma secretase inhibitor is N—[N-(3,5-Diflurophenaacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester (DAPT).
4. The method of claim 1 , further comprising transplanting the population of human PDX1-positive endoderm cells into a human host to mature into insulin secreting cells.
5. The method of claim 4 , wherein the insulin secreting cells form islet-like cell clusters.
6. The method of claim 5 , wherein the islet-like cell clusters comprise singly-positive insulin secreting cells.
7. The method of claim 1 , wherein the population of human PDX1-positive pancreatic endoderm cells expresses neurogenin (NGN3) and does not substantially express a marker selected from the group consisting of ghrelin, glucagon, insulin, and islet amyloid polypeptide (IAPP).
8. The method of claim 1 , further comprising contacting the population of human PDX1-positive pancreatic endoderm cells with a factor selected from the group consisting of nicotinamide (NIC), exendin 4 (Ex4), hepatocyte growth factor (HGF), insulin-like growth factor (IGF) and combinations thereof.
9. The method of claim 1 , wherein the retinoid is retinoic acid.