血管内皮生长因子基因体外转染大鼠神经干细胞的研究  

Gene transfer of vascular endothelial growth factor plasmid/liposome complexes in rat neural stem cells in vitro

作　　者：朱巍[1] 周良辅[1] 汪洋[2] 朱剑虹[1] 毛颖[1] 

机构地区：[1]复旦大学附属华山医院神经外科,上海200040 [2]复旦大学上海医学院解剖组胚系

出　　处：《中华实验外科杂志》2004年第6期730-732,i006,共4页Chinese Journal of Experimental Surgery

基　　金：上海市科技发展基金重点资助项目 (0 2DJ1 4 0 2 5)

摘　　要：目的　探讨经脂质体介导转染血管内皮生长因子 (VEGF) 12 1基因的大鼠胚胎神经干细胞体内外基因表达的特点。方法　脂质体介导法将VEGF12 1基因转染到大鼠神经干细胞中。经逆转录 聚合酶链反应 (RT PCR)及免疫荧光染色检测转基因神经干细胞及其分化细胞的基因表达情况。用立体定向法将BrdU标记的转基因神经移植到大鼠大脑中动脉梗死模型的纹状体缺血半暗区 ,免疫荧光染色观察移植后 1周转基因神经干细胞在脑内的基因表达情况。结果　转基因神经干细胞及其分化的子代细胞均有VEGF12 1的表达并持续 2周左右。转基因神经干细胞移植后 1周在宿主脑内迁徙并表达VEGF基因产物。结论　经脂质体介导转染VEGF12Objective To detect the gene expression of rat neural stem cells (NSCs) transfected with vascular endothelial growth factor (VEGF) mediated by lipofectamine.Methods VEGF121 gene was transferred to rat NSCs by using lipofectamine technique.Gene expression of transfected cells was detected by RT PCR and immunofluorescent staining in vitro.Temporary middle cerebral artery occlusion (tMCAO) models of rats were established.BrdU labelled VEGF121 transfected NSCs were transplanted into the penumbra zones 3 days after tMCAO model established.By using immunofluorescent staining,VEGF expression of transplanted cells 1 week after transplantation was detected.Results VEGF transfected NSCs could continuously express gene products during the first 2 weeks.Both transfected NSCs and their progeny expressed VEGF gene products,which was demonstrated by fluorescence study.One week after transplantation,immunofluorescent staining showed that VEGF transfected NSCs migrated and expressed VEGF in hosts' brains.Conclusion NSCs which are transfected with VEGF121 gene by lipofectamine can express gene products efficiently in vitro and in vivo.

关 键 词：血管内皮生长因子 大鼠 神经干细胞 基因转移 移植 

分 类 号：R346[医药卫生—基础医学]