US 12,168,028 B2
Methods for obtaining cardiomyogenic precursor cells
Joshua M. Hare, Miami Beach, FL (US); and Konstantinos Chatzistergos, Miami, FL (US)
Assigned to UNIVERSITY OF MIAMI, Miami, FL (US)
Appl. No. 16/649,522
Filed by UNIVERSITY OF MIAMI, Miami, FL (US)
PCT Filed Sep. 21, 2018, PCT No. PCT/US2018/052215
§ 371(c)(1), (2) Date Mar. 20, 2020,
PCT Pub. No. WO2019/060721, PCT Pub. Date Mar. 28, 2019.
Claims priority of provisional application 62/562,150, filed on Sep. 22, 2017.
Prior Publication US 2020/0246389 A1, Aug. 6, 2020
Int. Cl. A61K 35/34 (2015.01); A61K 38/25 (2006.01); C12N 1/38 (2006.01); C12N 5/074 (2010.01); C12N 5/077 (2010.01); C12N 5/10 (2006.01)
CPC A61K 35/34 (2013.01) [A61K 38/25 (2013.01); C12N 1/38 (2013.01); C12N 5/0657 (2013.01); C12N 5/0696 (2013.01); C12N 5/10 (2013.01); C12N 2501/415 (2013.01); C12N 2501/60 (2013.01); C12N 2501/85 (2013.01); C12N 2506/45 (2013.01)] 4 Claims
 
1. A method of obtaining a population of human cardiomyogenic precursor cells, the method comprising:
(a) differentiating human induced pluripotent stem cells (hiPSCs) in the presence of all of a glycogen synthase kinase 3 (GSK3) inhibitor, a Wnt inhibitor, and recombinant growth hormone releasing hormone or their synthetic analogs to obtain hiPSC-derived human cardiac precursor cells; and
(b) isolating a population of human cardiac precursor cells based on (i) expression of human growth hormone-releasing hormone receptor (GHRHR) using a GHRHR antibody and (ii) no expression of c-Kit to obtain a population consisting essentially of hiPSC-derived human cardiomyogenic precursor cells which express Nkx2.5 and Isl1.