Transfection of adeno-associated virus encoding beta-nerve growth factor into endothelial progenitor cells

doi:10.3969/j.issn.2095-4344.2015.36.021

Abstract

Abstract:

BACKGROUND: Nowadays, gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. Its key is to choose proper cells, genes and vectors.

OBJECTIVE: To use recombinant adeno-associated virus mediated β-nerve growth factor (β-NGF) to transfect rat bone marrow-derived endothelial progenitor cells in vitro, and to investigate the effect of β-NGF expression on the proliferation of endothelial progenitor cells.

METHODS: The endothelial progenitor cells were isolated, cultured and identified from the bone marrow of rats. Empty vector or recombinant adenovirus-associated virus containing β-NGF gene was transferred into endothelial progenitor cells. We examined the transfection efficiency by fluorescence expression of green fluorescent protein. Expression of β-NGF protein was detected using ELISA, and its effect on the proliferation of endothelial progenitor cells was determined using MTT method.

RESULTS AND CONCLUSION: Rat endothelial progenitor cells were isolated and cultured successfully in vitro and were identified positive by the function of cells and immunofluorescence staining. The endothelial progenitor cells were infected directly by the recombinant adenovirus-associated virus containing β-NGF gene with an efficiency of 65.3%. β-NGF protein was detected in the culture supernatant of transfected endothelial progenitor cells, which reached a high level at 10 days after gene transfection. Furthermore, there was no β-NGF protein in the blank and empty vector groups. After transfection, the proliferative ability of endothelial progenitor cells was increased, which was significantly higher than the blank and empty vector groups (P < 0.05). But there was no difference between the latter two groups (P > 0.05). These findings suggest that recombinant adenovirus-associated virus containing β-NGF gene can be successfully transferred into rat bone marrow-derived endothelial progenitor cells and promote the proliferation of endothelial progenitor 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

Key words: Nerve Growth Factor, Cells, Cultured, Genes, Transfection

CLC Number:

R394.2

Yang Zhong-yan, Gao Chun-zheng, Wu Dong-jin, Peng Chang-liang. Transfection of adeno-associated virus encoding beta-nerve growth factor into endothelial progenitor cells [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(36): 5853-5858.

Figures/Tables 3

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