[Effects of cardiotrophin-1 on differentiation of cardiomyocyte-like cells induced from rat bone marrow mesenchymal stem cells].
OBJECTIVE
To investigate the effects of cardiotrophin-1 (CT-1) on differentiation of induced rat bone marrow mesenchymal stem cells (BMMSCs) in vitro with 5-azacytidine (5-aza) for the purpose of elucidation of the cellular biological mechanisms.


METHODS
BMMSCs isolated from femur of rats were divided into four groups: untreated group as control (group A); 0.1 nmol/L CT-1 added to medium (Group B); induced with 10 micromol/L 5-aza only (Group C); induced with 10 micromol/L 5-aza combined with 0.1 nmol/L CT-1 added to medium (Group D). After 4 weeks of induced culturing, the differentiation of induced myocyte like cells were estimated, levels of cardiac troponin-T (cTnT) by immunohistochemical staining and ultrastructure of induce-cultured BMMSCs were determined and mRNA expression of alpha-actin, beta-myosin heavy chain (beta-MHC), Nkx2.5, GATA4 were analyzed by real time polymerase chain reaction (RT-PCR).


RESULTS
After 4 weeks of induced culturing, morphological characteristics of myocyte like cells with the expression of cTnT were observed in group C and D cells. Higher level expression of GATA4, Nkx2.5, alpha- actin and beta-MHC mRNA in group D was observed by comparing with those of group C and the differentiated BMMSCs with formations of myofilaments distinctly were also existed in 5-aza combined with CT-1 treatment group.


CONCLUSIONS
This study suggests that induced culturing of BMMSCs in the presence of 5-aza combined with CT-1 can enhance cardiomyocytic characteristics. CT-1 upregulates the expression of GATA4, Nkx2.5, alpha-actin and beta-MHC mRNA, and rapidly promotes the differentiation and maturation of cardiomyocyte-like cells differentiated from BMMSCs induced with 5-aza.
Chen
Xin-Yun
XY
Department of Cardiology, West China Hospital, Sichuan University, Chengdu 610041, China.
Zeng
Zhi
Z
Rao
Li
L
Li
Sheng-Fu
SF
Zhou
Bin
B
Chen
Yu-Cheng
YC
D000818:Animals; D001374:Azacitidine / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D003470:Culture Media; D016207:Cytokines / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D003470:Culture Media; D016207:Cytokines; C092073:cardiotrophin 1; D001374:Azacitidine