Effect of Let-7c on neural differentiation of bone marrow mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2016.01.004

Abstract

Abstract:

BACKGROUND: The microRNAs are involved in regulation of stem cell proliferation, differentiation and aging. To study the effect of Let-7c, a member of Let-7, on the neural differentiation of bone marrow mesenchymal stem cells provides new ideas for stem cell therapy.

OBJECTIVE: To investigate the role of Let-7c in the neural differentiation of bone marrow mesenchymal stem cells.

METHODS: The lentiviral vectors of Let-7c-up and Let-7c-inhibition were constructed and transfected into rat bone marrow mesenchymal stem cells. Optimal multiplicity of infection was screened. The cells were divided into non-transfected group, negative control group (transfected with empty virus), transfected enhancement group (transfected with LV-rno-Let-7c-up), transfected inhibition group (transfected with LV-rno-Let-7c-5p-inhibition). Bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The fluorescence expressed by transfected cells was observed under inverted fluorescence microscope. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The cell viability was determined by MTT method.

RESULTS AND CONCLUSION: Under the inverted fluorescence microscope, the cells were successfully transfected with LV-rno-Let-7c-up and LV-rno-Let-7c-5p-inhibition. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in the transfected enhancement group were significantly higher than those in the negative control group (P < 0.05), while in the transfected inhibition group, they were lower than those in the negative control group (P < 0.05). These findings indicate that the differentiation percentage of bone marrow mesenchymal stem cells is increased by fasudil after transfection with LV-rno-Let-7c-up, and Let-7c may promote the differentiation of bone marrow mesenchymal stem cells into neurons.

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

CLC Number:

R394.2

Wang Jing, Zhao Shao-yun, Li Ming-zhe, Jing Li-jun, Jiao Shu-jie, Peng Tao, Teng Jun-fang, Jia Yan-jie . Effect of Let-7c on neural differentiation of bone marrow mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(1): 20-25.

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