Journal of Shanghai University(Natural Science Edition) ›› 2017, Vol. 23 ›› Issue (3): 378-386.doi: 10.12066/j.issn.1007-2861.1939

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Human embryonic stem cell-derived cardiomyocytes model establishment and systemic identification scheme

CHEN Xinxin, LIU Zhuyuan, GU Huanyu, ZHOU Lei   

  1. Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2017-04-03 Online:2017-06-30 Published:2017-06-30

Abstract:

Cardiomyocytes is useful in the study of many cardiovascular diseases, while human cardiomyocytes are difficult to obtain and culture. With multi-directional differentiation properties, human embryonic stem cells can provide cell resources in vitro. This study aims to establish a useful protocol to induce the differentiation of human embryonic stem cells into cardiomyocytes, and observe identification of human cardiomyocytes. Expression of stem cell markers including Cripto, Dnmt3b, Wnt3, KIF4, Oct4, SOX2 and Nanog decrease during differentiation while cardiac progenitor cell marker Nkx2.5 and cardiomyocytes specific markers TnnT2 and a-actinin increase. Ratios of TnnT2 positive cells, α-actinin positive cells, TnnT2 and α-actinin double positive cells are 90.80%, 91.00%, and 90.91%. These results indicate that human embryonic stem cells can be efficiently induced into cardiomyocytes in the proposed protocol. In conclusion, differentiation of human embryonic stem cells into cardiomyocytes has been successfully induced and an identification scheme has been established, and markers have been detected through quantitative PCR and immunofluorescence stainings. The protocol will facilitate better understanding
of pathogenesis of cardiovascular diseases and enable better cardiotoxicity drug screening.

Key words: cardiomyocytes, cell differentiation, human embryonic stem cell