摘要
目的 :检测逆转录病毒介导血管内皮细胞生长因子(VEGF1 65)基因转染兔骨髓基质干细胞 (BMSC)后目的基因表达情况及促血管生成作用 ,为进一步将其用于血管化组织工程骨构建奠定基础 .方法 :构建含VEGF基因的逆转录病毒表达载体 ,通过感染兔BMSC 使其获得稳定表达 ,使用免疫组化的方法检测其蛋白表达 ,并通过新生血管计数观察其促血管生成作用 .结果 :成功构建VEGF逆转录病毒表达载体 ,免疫组化方法显示经感染的BMSC 胞质中有阳性棕色颗粒出现 ,且动物实验显示新生血管计数明显高于对照组 (P <0 .0 1 ) .结论 :采用逆转录病毒介导的基因转移技术可将VEGF基因转染至BMSC 中 ,可获得外源性蛋白的表达 ,且有明显的促血管生成作用 .
AIM: To detect the expression of retrovirus transfected vascular endothelial growth factor (VEGF) in rabbit mesenchymal stem cells and its effect on angiogenesis, to lay the foundation for its further use in vascularized tissue engineered bone. METHODS: A retroviral vector containing VEGF gene was constructed and expressed in BMS C. The protein expression was detected by immunohistochemistry and its effect of stimulating angiogenesis was observed by microvessel count (MVC). RESULTS: Retroviral vector containing VEGF gene was constructed successfully. Immunohistochemistry showed that it was expressed in BMS C plasma (brown staining). MVC in experimental group was higher than that in control group ( P < 0.01). CONCLUSION: Gene transfer technology mediated by retroviral vector can transfect VEGF gene into BMS C and obtain the expression of exogenous protein which has obvious effect of stimulating angiogenesis.
出处
《第四军医大学学报》
北大核心
2003年第24期2262-2265,共4页
Journal of the Fourth Military Medical University
