鼠IGF-1基因转染鼠神经干细胞的研究  

Experimental study on insulin-like growth factor-1 gene transfecting neural stem cells in vitro

作　　者：张旭[1] 藏卫东[1] 胡婵丽[1] 

机构地区：[1]郑州大学基础医学院生物化学教研室,郑州450052

出　　处：《中国卫生检验杂志》2006年第3期357-359,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的:构建胰岛素样生长因子-1(IGF-1)基因修饰的神经干细胞(NSCs),用于神经系统疾病的治疗。方法:通过酶切和连接的方法构建IGF-1基因重组逆转录病毒载体pLXSN-IGF-1,转染包装细胞获得病毒液,测滴度后感染从胎龄14 d的SD大鼠全脑中分离出的神经干细胞(NSCs),在转基因NSC中检测IGF-1基因的表达,观察转基因对NSCs增殖的影响。结果:酶切及序列分析证实逆转录病毒载体pLXSN-IGF-1构建正确,NSCs转基因后能检测到IGF-1基因表达,且仍然保持着未分化状态,能够自我更新以及具有多向分化潜能,并且生长速率加快。结论:成功构建了转IGF-1基因神经干细胞。Objective： To construct transgenic neural stem cells containing IGF - 1 gene for therapy of nervous system diseases. Methods：The recombinant retroviral vector containing IGF - 1 gene pLXSN - IGF - 1 was established using the method of enzyme cutting and ligation. The recombinant vector was transferred to packaging cell line - pA317 to acquair virus. After detection for titration, the recombinant virus was used to infect neural stem cells （NSCs） isolated from the whole brain of embryonic day 14 （E14） SD rats. We detected the expression of IGF - 1 gene and observed the effect of transgene on proliferation of NSCs. Results：The retroviral vector pLXSN -IGF- 1 was successfully established, and it was proved by enzyme cutting and sequence analysis. The expression of IGF - 1 gene was confirmed. NSC transfected with IGF - 1 gene remained undifferentiated and had self- renewing capacity and potential multipotency of differentiation. NSC transfected by IGF - 1 gene grew faster than that free from transfection. Conclusion：The IGF- 1 transgene NSCs are successfully constructed.

关 键 词：神经干细胞 IGF-1 逆转录病毒 转染 转基因 

分 类 号：Q785[生物学—分子生物学]