LncRNA-Braveheart promotes the differentiation of bone marrow mesenchymal stem cells in vitro into cardiomyocyte-like cells

doi:10.3969/j.issn.2095-4344.2017.29.001

Abstract

Abstract:

BACKGROUND: Our previous work demonstrated that bone marrow mesenchymal stem cells (BMSCs) transplantation could improve cardiac function in rats with myocardial infarction. However, the overall efficacy was unsatisfactory, and there was a low efficiency of BMSCs differentiating into cardiomyocytes in the local infarct myocardium.
OBJECTIVE: To transfect long non-coding RNA-Braveheart (lncRNA-Bvht) into BMSCs in order to observe whether it could promote cardiomyocyte differentiation of BMSCs in vitro.
METHODS: pLVX-IRES-ZsGreen1-lncRNA-Bvht vector was constructed and applied to transfect lncRNA-Bvht into bMSCs, and then, the transfection efficiency was detected. BMSCs were obtained from C57BL/6 mice and cultured in vitro. Passage 3 cells were divided into three groups: BMSCs group, null vector group and lncRNA-Bvht group. All cells in the three groups were cultured in the normal condition for 48 hours and cardiomyocytes differentiation was induced by 5-azacytidine for another 24 hours followed by 2-week culture under normal conditions. Cardiomyocyte differentiation of BMSCs was observed under fluorescence microscopy and expression of cardiac specific cell markers including troponin T and myosin were examined using immunofluorescent staining, western blot assay, and qRT-PCR.
RESULTS AND CONCLUSION: Cell morphological changes could be observed in all the groups 2 weeks after the induction. Interconnected cells arranged consistently in all the three groups. Immunofluorescent staining results showed that the expression of troponin T and myosin was notably positive, and the proportion of troponin T positive cells was significantly increased. qRT-PCR and western blot assay results indicated that there were significantly increased levels of troponin T and myosin in the lncRNA-Bvht group as compared with the BMSCs and null vector groups (P < 0.01), suggesting that lncRNA-Bvht could efficiently promote cardiomyocyte differentiation of BMSCs in vitro.

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

Key words: Myocardial Infarction, Bone Marrow, Mesenchymal Stem Cells, Myocytes, Cardiac, Tissue Engineering

CLC Number:

R394.2

Hou Jing-ying, Long Hui-bao, Zhou Chang-qing, Wu Hao, Guo Tian-zhu, Zhong Ting-ting, Wu Quan-hua, Wang Lei, Zheng Shao-xin, Wang Tong. LncRNA-Braveheart promotes the differentiation of bone marrow mesenchymal stem cells in vitro into cardiomyocyte-like cells[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(29): 4593-4599.

Figures/Tables 2

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