Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (19): 2981-2986.doi: 10.3969/j.issn.2095-4344.2014.19.005

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Differentiation of mesenchymal stem cells into cardiomyocyte-like cells induced by H9C2 cell culture medium

Huang Lv, Zhang Hui, Xie Li, Li Yong-lin, Zhang Xiao-gang   

  1. Department of Cardiology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Revised:2014-04-07 Online:2014-05-07 Published:2014-05-07
  • Contact: Zhang Hui, Physician, Department of Cardiology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • About author:Huang Lv, Studying for master’s degree, Department of Cardiology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Supported by:

    the Medical Science and Technology Project of Chongqing Health Bureau, No. 2009-1-51

Abstract:

BACKGROUND: Mesenchymal stem cells can differentiate into cardiomyocytes in vitro induced by different methods, such as chemical drug induction, autologous transplantation in vivo, and in vitro simulation of cardiac-like microenvironment, but these inducible methods show low induction rate, complex operation, and toxic side effects.
OBJECTIVE: To investigate the role of H9C2 cell culture medium in the differentiation of mesenchymal stem cells into cardiomyocyte-like cells.
METHODS: Mesenchymal stem cells were obtained by the whole bone marrow adherent culture and H9C2 cell culture medium was prepared as a culture medium. Then mesenchymal stem cells were co-cultured with H9C2 cell culture medium for 1, 3, 5, 7 days. H9C2 cells cultured in 10% Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 served as positive control groups, while mesenchymal stem cells cultured in 10% Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 as negative control group. Immunofluorescence and western blot assay were used to detect expression of myocardial cell junction protein (desmin) and troponin T. Real-time quantitative PCR was applied to detect mRNA expression of myocardial cell trait genes, α-cardiac myosin heavy chain and β-myosin heavy chain.
RESULTS AND CONCLUSION: After co-culture of H9C2 cell culture medium and mesenchymal stem cells for 7 days, the troponin T positive cells were up to (16±7)%, which was significantly higher than that of non-induced mesenchymal stem cells. Compared with the negative control group, the expression of troponin T protein and desmin after induction were significantly increased (P < 0.05) by western-blot detection; real-time PCR showed that the mRNA expression of α-cardiac myosin heavy chain and β-myosin heavy chain in differentiated cells were both up-regulated (P < 0.05). These findings suggest that H9C2 cell culture medium may induce the differentiation of mesenchymal stem cells into cardiomyocyte-like cells.


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


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Key words: mesenchymal stem cells, myocytes, cardiac, troponin T

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