Effects of transfection with acidic fibroblast growth factor by electroporation on skeletal muscle satellite cells

doi:10.3969/j.issn.2095-4344.2015.01.015

Abstract

Abstract:

BACKGROUND: Previous studies have shown that a certain dose of acidic fibroblast growth factor can promote skeletal muscle satellite cell proliferation in vitro.
OBJECTIVE: To investigate the effects of transfection with acidic fibroblast growth factor by electroporation on growth, proliferation and differentiation of skeletal muscle satellite cells.
METHODS: Skeletal muscle satellite cells were cultured and purified, and then transfected with plasmid pSectag-GFP-aFGF by electroporation. The expression of green fluorescent protein was observed under fluorescence microscope, and the transfection efficiency was calculated. After transfection, cell cycle was analyzed by flow cytometry to draw the growth curve of skeletal muscle satellite cells. Western blot assay was employed to measure protein level of acidic fibroblast growth factor.
RESULTS AND CONCLUSION: (1) Immunocytochemistry detection: The skeletal muscle satellite cells were positive for a-sarcomeric actin. (2) Transfection efficiency: At 12 hours after transfection with pSectag-aFGF, several cells showed green fluorescence, and the green fluorescent expression reached the peak at 72-96 hours after transfection with a positive rate of about 90%. (3) Cell cycle: After electrotransfection, the proportion of cells at S phase in the electroporation group was higher than that in the control group (P < 0.05). (4) Cell growth curve: At 3 days after electrotransfection, the cells entered logarithmic growth phase but the proliferation slowed down at 5 days. (5) Differentiation capacity: There were fewer myotubes and aging cells in the electroporation group than the control group. (6) Western blot assay: Acidic fibroblast growth factor protein was highly expressed in the cells transfected with target gene detected by western blot assay. These findings indicate that by using electroporation method, acidic fibroblast growth factor can be transferred into skeletal muscle satellite cells and have a high-efficiency and long-term expression, which can promote the proliferation of skeletal muscle satellite cells and inhibit formation of myotubes.

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

全文链接：

Key words: Satellite Cells, Skeletal Muscle, Genes, Transfection, Green Fluorescent Proteins

CLC Number:

R394.2

Li Jiang-hua, Dong Shao-hong, Xiong Wei, Pang Xin-li, Liu Qi-yun, Li Wen-jun . Effects of transfection with acidic fibroblast growth factor by electroporation on skeletal muscle satellite cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(1): 85-90.

Figures/Tables 2

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