中国组织工程研究与临床康复2011，Vol.15Issue(10)：1769-1772,4.DOI:10.3969/j.issn.1673-8225.2011.10.013
人血管内皮生长因子165基因转染兔骨髓间充质干细胞的蛋白表达
Protein expression of bone marrow mesenchymal stem cells after human vascular endothelial growth factor 165 gene transfection
摘要
Abstract
BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) is the best source of seed cells for tissue engineering, which containing vascular endothelial growth factor 165 (VEGF165) not only has an important significance in angiogenesis and the start of bone repair, but also its sustained release can increase the degree of mineralization of new bone, and enhance the mechanical properties of repair tissue.OBJECTIVE: To observe the hVEGF165 protein function expressed by the gene transfected BMSCs.METHODS: BMSCs from rabbits were isolated and cultured in vitro, and which were purified and identified. Cell surface makers were detected by immunofluorescence assay. And the cultured BMSCs were transfected with the composite of PcDNA3.1-hVEGF165 plasmid and lipofectamine in the ratio of 1/3 mixed liquor, and which were divided into 3 groups:transfection group, empty vector transfection group, and non-transfection group. PcDNA3.1-hVEGF165 plasmid was used to transfect cells in transfection group; pcDNA3.1-empty vector transfection was applied in empty vector transfection group; no treatment was performed in non-transfection group. The expression of exogenous hVEGF165 was detected by ELISA and Western-blot.RESULTS AND CONCLUSION: Compared with empty vector transfection group and non-transfection g roup, the protein level of VEGF165 in transfection group was significantly higher (P ＜ 0.05), however, there was no significant difference between empty vector transfection group and non-transfection group (P ＞ 0.05). There were significant differences of the protein level of VEGF165 in transfection group (P ＜ 0.05) at different time points. The transfected BMSCs by hVEGF165 can successfully secret the protein of VEGF165. It is indicated that hVEGF165 can be transfected to BMSCs and efficiently express VEGF165 protein of biological activity by using of gene transfection technology .关键词
血管内皮生长因子165/基因转染/骨髓间充质干细胞/质粒/蛋白分类
基础医学引用本文复制引用
田力,梁晓鹏,田晓晔,于鑫琛,田晶..人血管内皮生长因子165基因转染兔骨髓间充质干细胞的蛋白表达[J].中国组织工程研究与临床康复,2011,15(10):1769-1772,4.基金项目
辽宁省教育厅基金资助项目(2008-7-29) （2008-7-29）
沈阳科技局基金资助项目(1081233-1-00-03). （1081233-1-00-03）
