胶质细胞源性营养因子及内皮素B受体基因共转染神经干细胞实验研究  

Study of the cotransfection of GDNF and EDNRB into neural stem cells

作　　者：陈景波[1] 王国斌[1] 孙念峰[1] 陶凯雄[1] 舒晓刚[1] 张景辉[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院腔镜外科,湖北武汉430030

出　　处：《中国现代医学杂志》2009年第10期1465-1468,1473,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(No:30371397)

摘　　要：目的探讨神经干细胞转染的新方法,并观察转染后目的基因的表达情况。方法原代培养神经干细胞,运用jetPEI转染试剂将目的基因胶质细胞源性神经营养因子(GDNF)和内皮素B受体基因(EDNRB基因)共转染至神经干细胞内,免疫荧光显微镜观察、流式细胞仪检测绿色荧光蛋白(GFP)表达情况,测定转染效率,RT-PCR检测目的基因表达情况。结果成功培养出可用于基因转染的神经干细胞,转染后24h即可在免疫荧光显微镜下观察到GFP表达,流式细胞仪显示24、48和72h转染效率分别为15.36%、24.67%和25.73%。RT-PCR显示目的基因在神经干细胞内成功表达。结论运用jetPEI成功将目的基因转染至神经干细胞内,为相关神经变性疾病的基因治疗打下了实验基础。[Objectives] To investigate the optimizing method of neural stem eel/（NSC） transfection and the expression of extrinsic genes in neural stem cells. [Methods] Extrinsie genes GDNF and EDNRB were eotransfeeted into primary Cultured neural stem ceils by using jet PEI. The expression of green fluorescent protein （GFP） was measured with fluorescence microscope and flow eytometer. To detect the expression of mRNA of GDNF and EDNRB by RT-PCR. [Results] Bright green fluorescence of the transfected eells could be observed under fluorescence mieroscope after 24 h of transfeetion. Flow cytometer analysis showed that the effieieney of eotransfeetion were 15.36%, 24.67%, 25.73% in 24, 48 and 72 hours respectively. Meanwhile the expression of GDNF and EDNRB genes were detected in neural stem ceils by RT-PCR. [Conclusions] The target genes were successfully eotransfeeted into neural stem cells by using jet PEI. It provides a feasible technological platform for the polygene therapy of neural degenerative diseases.

关 键 词：GDNF基因 EDNRB基因 神经干细胞 细胞转染 

分 类 号：R781[医药卫生—口腔医学]