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Experimental research
Effects of cardiotrophin-1 on differentiation and maturation of rat bone marrow mesenchymal stem cells induced with 5-azacytidine in vitro
  1. Chen Xinyun1,
  2. Zeng Zhi1,
  3. Zhou Bin2,
  4. Rao Li1,
  5. Chen Yucheng1,
  6. Yan Yafei3,
  7. Zhang Tingjie3,
  8. Li Shengfu4
  1. 1Department of Cardiology, West China Hospital, Sichuan University, Chengdu, China
  2. 2Laboratory of Molecular Translational Medicine, West China Second University Hospital, Sichuan University, Chengdu, China
  3. 3Department of Cardiology, The No.1 municipal Hospital of Chendu, Chengdu, China
  4. 4Key Laboratory of Transplant Engineering and Immunology of Ministry of Health, West China Hospital, Sichuan University, Chengdu, China

Abstract

Background Cardiotrophin-1 (CT-1) is a cytokine involved in the growth and survival of cardiac cells that stimulates cardiomyogenesis in pluripotent murine embryonic stem (ES) cells. But it is not known whether CT-1 is responsible for the fate of differentiated bone marrow mesenchymal stem cells (BMMSCs).

Methods We investigated the effects of CT-1 on differentiation and maturation of BMMSCs in vitro induced with 5-azacytidine. After 4 weeks of induced culturing, we observed the levels of α-cardiac actin and troponin-I by immunohistochemical staining, the ultrastructure of induce-cultured BMMSCs, and the expression of GATA-4, NKx2.5, beta-myosin heavy chain (beta-MHC) and α-cardiac actin mRNA by real time RT-PCR analysis.

Results Differentiated BMMSCs cultured with CT-1 distinctly showed formations of myofilaments-morphological characteristics of myocyte like cells. The protein levels of α-cardiac actin and troponin-T were significantly higher than control. Furthermore, mRNA expression of GATA-4, NKx2.5, α-cardiac actin and beta-MHC was increased remarkably.

Conclusions This study suggests that induced culturing of BMMSCs in the presence of 5-azacytidine combined with CT-1 can enhance cardiomyocytic characteristics. CT-1 upregulates the expression of GATA-4, NKx2-5, α-cardiac actin and beta-MHC mRNA, and rapidly promotes the differentiation and maturation of cardiomyocyte-like cells differentiated from BMMSCs induced with 5-azacytidine.

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