Effects of cardiotrophin-1 on differentiation and maturation of rat bone marrow mesenchymal stem cells induced with 5-azacytidine in vitro

Xinyun, Chen; Zhi, Zeng; Bin, Zhou; Li, Rao; Yucheng, Chen; Yafei, Yan; Tingjie, Zhang; Shengfu, Li

doi:10.1016/j.ijcard.2009.02.009

« Previous Next »International Journal of Cardiology
Volume 143, Issue 2 , Pages 171-177, 20 August 2010

Effects of cardiotrophin-1 on differentiation and maturation of rat bone marrow mesenchymal stem cells induced with 5-azacytidine in vitro

Chen Xinyun
- Department of Cardiology, West China Hospital, Sichuan University, Chengdu, 610041, China
- Department of Cardiology, The No.1 municipal Hospital of Chendu, Chengdu 610041, China
Zeng Zhi
- Department of Cardiology, West China Hospital, Sichuan University, Chengdu, 610041, China
- Corresponding author. Tel.: +86 28 15828179639; fax: +86 28 85315539.
Zhou Bin
- Laboratory of Molecular Translational Medicine, West China Second University Hospital, Sichuan University, Chengdu 610041, China
Rao Li
- Department of Cardiology, West China Hospital, Sichuan University, Chengdu, 610041, China
Chen Yucheng
- Department of Cardiology, West China Hospital, Sichuan University, Chengdu, 610041, China
Yan Yafei
- Department of Cardiology, The No.1 municipal Hospital of Chendu, Chengdu 610041, China
Zhang Tingjie
- Department of Cardiology, The No.1 municipal Hospital of Chendu, Chengdu 610041, China
Li Shengfu
- Key Laboratory of Transplant Engineering and Immunology of Ministry of Health, West China Hospital, Sichuan University, Chengdu, 610041, China

Received 9 October 2008; received in revised form 9 December 2008; accepted 6 February 2009. published online 09 March 2009.

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. BMMSCs isolated from femur of rats were induced by CT-1 only, by 5-azacytidine with or without CT-1,and an untreated control group was also set. After 4 weeks of induced culturing, we observed the levels of alpha-cardiac actin and troponin-I by immunohistochemical staining, the ultrastructure of induce-cultured BMMSCs, and the expression of GATA4, Nkx2.5, beta-myosin heavy chain (beta-MHC) and alpha-cardiac actin mRNA by real time RT-PCR analysis.

Results

Differentiated BMMSCs treated by 5-azacytidine and CT-1 distinctly showed formations of myofilaments and myotube-like structures—morphological characteristics of myocyte like cells, and spontaneous contraction of a few cells was observed. The protein levels of alpha-cardiac actin and troponin-T were significantly higher than control. Furthermore, mRNA expression of GATA4, Nkx2.5, alpha-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 GATA4, Nkx2–5, alpha-cardiac actin and beta-MHC mRNA, and rapidly promotes the differentiation and maturation of cardiomyocyte-like cells differentiated from BMMSCs induced with 5-azacytidine.

Keywords: Bone marrow mesenchymal stem cells, Cardiomyocyte-like cells, Cardiotrophin-1, 5-Azacytidine