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

• Published in: International journal of cardiology Vol. 143; no. 2; pp. 171 - 177
• Main Authors: Xinyun, Chen, Zhi, Zeng, Bin, Zhou, Li, Rao, Yucheng, Chen, Yafei, Yan, Tingjie, Zhang, Shengfu, Li
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 20-08-2010; Elsevier
• Subjects: 5-Azacytidine; Actins - genetics; Animals; Azacitidine - pharmacology; Biological and medical sciences; Bone Marrow Cells - cytology; Bone Marrow Cells - drug effects; Bone marrow mesenchymal stem cells; Cardiology. Vascular system; Cardiomyocyte-like cells; Cardiotrophin-1; Cardiovascular; Cell Differentiation - drug effects; Cell Survival - drug effects; Cells, Cultured; Cytokines - pharmacology; Enzyme Inhibitors - pharmacology; GATA4 Transcription Factor - genetics; Gene Expression - drug effects; Homeobox Protein Nkx-2.5; Homeodomain Proteins - genetics; In Vitro Techniques; Male; Medical sciences; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - drug effects; Myosin Heavy Chains - genetics; Rats; Rats, Sprague-Dawley; Transcription Factors - genetics; Cardiotrophin-1; Bone marrow mesenchymal stem cells; Cardiomyocyte-like cells; 5-Azacytidine; Heart; Rat; Stem cell; Rodentia; Cardiovascular disease; In vitro; Myocyte; Vertebrata; Mammalia; Azacitidine; Animal; Bone marrow; Cardiology; Differentiation
• ISSN: 0167-5273; 1874-1754
• DOI: 10.1016/j.ijcard.2009.02.009

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.