Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (19): 3431-3435.doi: 10.3969/j.issn.1673-8225.2010.19.003

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Differentiation of rabbit bone marrow mesenchymal stem cells into cardiomyocytes in vitro following induction: Changes in connexin 43 expressionv

Zhou Hao-yue1, Qiu Han-ying2, Lu Jiong-bin1，Li Xie-dong1   

  1. 1Medical College, Shantou University, Shantou   515000, Guangdong Province, China;
    2Department of Cardiology, Second Affiliated Hospital, Medical College, Shantou University, Shantou   515000, Guangdong Province, China
  • Online:2010-05-07 Published:2010-05-07
  • Contact: Qiu Han-ying, Doctor, Professor, Master’s supervisor, Department of Cardiology, Second Affiliated Hospital, Medical College, Shantou University, Shantou 515000, Guangdong Province, China hy8720@yeah.net
  • About author:Zhou Hao-yue, Studying for master’s degree, Medical College, Shantou University, Shantou 515000, Guangdong Province, China zhy2221@hotmail.com

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Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) could differentiate to cardiomyocytes-like cells and express connexin 43 (CX43) after induction by inducing factors. Cell transplantation could improve cardiac function or repair damaged cardiac pacing conduction system. While CX43 plays an important role in maintaining normal function of the heart.
OBJECTIVE: To observe CX43 expression in the cardiomyocytes-like cells which were induced from the rabbit BMSCs in vitro, and study the gap junction intercellular communication between BMSCs and cardionmyoctes following induction.
METHODS: Rabbit BMSCs were derived and cultured by Percoll discontinuous density gradient centrifugation and adherent culture method. Cells in the normal group did not receive any intervention. Cells in the induction group were induced by 5-azacitidine to differentiate into cardiomyocytes-like cells. Immunofluorescence and flow cytometry were used to determine CX43 expression in BMSCs before and after induction. Neonatal rat cardiomyocytes following primary culture of 1 day were seeded on above-mentioned two groups of cell plates. Immunofluorescence was used to study the gap junction between BMSCs and cardiomyocytes. Normal, induction and additional enoxolone blocker groups were established in cell scratch test to investigate the GJIC transformation before and after BMSC induction.
RESULTS AND CONCLUSION: CX43 had weak expression in the normal group, but the CX43 expression was significantly increased at 2 and 4 weeks following 5-azacitidine induction (P < 0.01). GJIC was significantly enhanced (P < 0.001), and showed dependent increase with prolonged induction time (P < 0.05). Following coculture of BMSCs and cardiomyocytes, CX43 expression showed significantly linear expression on contact surface of two neighboring cells in the induction group. Compared with that at 4 weeks following induction, GJIC was significantly suppressed in the blocker group (P < 0.001). Above-mentioned results indicated that BMSCs could express CX43 spontaneously, which showed gap junction with cardiomyocytes in the early transplantation, resulting in cardiac electrical conduction following transplantation and the paracrine effect of BMSCs.

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