Basic & Clinical Medicine ›› 2015, Vol. 35 ›› Issue (4): 463-469.

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Phorbol myristate acetate promotes differentiation of mouse induced pluripotent stem cells into cardiomyocytes in vitro

  

  • Received:2014-10-23 Revised:2014-12-30 Online:2015-04-05 Published:2015-04-08

Abstract: Objective To investigate the effect of PMA on the differentiation of mouse induced pluripotent stem cells into cardiomyocytes in vitro, and to establish an efficient and safe protocol for cardiac differentiation in vitro. Methods In this experiment, embryoid bodies (EBs) were formed by classical hanging drop method, and were directional differentiated to cardiomyocyte with PMA treatment; Furthermore, expression of cardiac troponin T and α-Sarcomeric actinin was observed by immunofluorescent staining, mRNA expression levels of the related genes Brachyury, cTnT, MLC2a, NKX2.5, GATA4 were analyzed by qPCR and RT-PCR.Control group was treated by DMSO, number of beating embryoid bodies was calculated. Results The best concentration of PMA to induce the cardiac differentiation of miPSC was 100 nmol/L and the beating area was found in 53% of embryoid bodies, compared with the control group (beating area was found in 12% of embryoid bodies), Differentiation ef?ciency was higher in PMA treatment group. IPS-CM expressed serve cardic related proteins and genes, and had the Structure characteristics of myocardium cell. Conclusion: PMA can promote miPSC to differentiate into cardiomyocyte in vitro.

Key words: iPSC, cardiomyocyte, PMA, differentiate