US 12,435,315 B2
Generating dorsal foregut, and anterior domain, endoderm cells
Jan Jensen, Cleveland, OH (US); and Michael A. Bukys, Cleveland, OH (US)
Assigned to The Cleveland Clinic Foundation, Cleveland, OH (US)
Appl. No. 17/609,954
Filed by The Cleveland Clinic Foundation, Cleveland, OH (US)
PCT Filed May 22, 2020, PCT No. PCT/US2020/034201
§ 371(c)(1), (2) Date Nov. 9, 2021,
PCT Pub. No. WO2020/237141, PCT Pub. Date Nov. 26, 2020.
Claims priority of provisional application 62/851,348, filed on May 22, 2019.
Prior Publication US 2022/0220447 A1, Jul. 14, 2022
Int. Cl. C12N 5/071 (2010.01); C07K 14/51 (2006.01); A61K 38/00 (2006.01)
CPC C12N 5/0678 (2013.01) [C07K 14/51 (2013.01); A61K 38/00 (2013.01); C12N 2501/155 (2013.01); C12N 2501/385 (2013.01); C12N 2506/02 (2013.01); C12N 2506/45 (2013.01)] 20 Claims
 
1. A method of generating pancreatic endoderm cells having dorsal pancreatic identity comprising:
a) contacting a population of pluripotent stem cells with a retinoic acid signaling pathway agonist and a bone morphogenetic (BMP) pathway inhibitor;
b) culturing at least a portion of said population of pluripotent stem cells such that a population of dorsal foregut endoderm (DFE) cells is generated;
wherein said stem cells are not exposed to a transforming growth factor beta (TGFB) pathway agonist during said culturing in b) or during said contacting in a);
c) contacting at least a portion of said population of DFE cells with a retinoic acid signaling pathway agonist and a FGFR pathway inhibitor; and
d) culturing a least a portion of said population of DFE cells such that a population of pancreatic endoderm (PE) cells having dorsal pancreatic identity is generated; and
wherein said PE cell are not exposed to a TGFB pathway agonist during said culturing in step d) or during said contacting in step c).