US 11,788,060 B2
Small molecule compound composition that efficiently induces differentiation of human pluripotent stem cells into myocardial cells
Qing Jing, Shanghai (CN); Xiaoxu Qui, Shanghai (CN); Yang Liu, Shanghai (CN); and Yifan Zhang, Shanghai (CN)
Assigned to Shanghai Institutes for Biological Sciences Chinese Academy of Sciences, Shanghai (CN)
Appl. No. 16/71,599
Filed by Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, Shanghai (CN)
PCT Filed Jan. 12, 2017, PCT No. PCT/CN2017/070989
§ 371(c)(1), (2) Date Jul. 20, 2018,
PCT Pub. No. WO2017/124963, PCT Pub. Date Jul. 27, 2017.
Claims priority of application No. 2016100381160.4 (CN), filed on Jan. 20, 2016.
Prior Publication US 2019/0032016 A1, Jan. 31, 2019
Int. Cl. C12N 5/077 (2010.01); A61P 9/00 (2006.01); C12N 5/0735 (2010.01); C12N 5/0789 (2010.01)
CPC C12N 5/0655 (2013.01) [A61P 9/00 (2018.01); C12N 5/0606 (2013.01); C12N 5/0647 (2013.01); C12N 5/0657 (2013.01); C12N 2501/04 (2013.01); C12N 2501/415 (2013.01); C12N 2501/70 (2013.01); C12N 2506/45 (2013.01)] 6 Claims
 
1. A method for producing a cardiomyocyte, wherein said method comprises (1) culturing human pluripotent stem cells on plates coated with a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma and in human pluripotent stem cell culture medium for 2 days, (2) after the 2 days, supplementing the medium of (1) with 12 μM CHIR plus 10 nM rapamycin and wherein the cells are cultured for 3 days, (3) after the 3 days, transferring and then culturing the cells of (2) in a medium comprising RPMI/B27 minus insulin supplemented with 12 μM CHIR plus 10 nM rapamycin without insulin for 1 day, and (4) after the 1 day, supplementing the medium of (3) with XAV939 plus 10 μM KY02111 for 4 days without medium replacement.