HIV-1来源的慢病毒载体转染大鼠脊髓神经干细胞

Lentivirus vector deriving from human immunodeficiency virus-1 transfer to rat spinal cord neural stem cells

目的介绍人类免疫缺陷病毒(Human immunodeficieney virus-1,HIV-1)载体的构建及转染大鼠脊髓神经干细胞的实验方法。方法将含有绿色荧光蛋白(green fluorescence protein,GFP)的4种质粒通过293T细胞构建成GFP-HIV载体。用Elisa法及Hela细胞分别验证其高浓度及强感染力后将其转染大鼠脊髓神经干细胞。结果GFP-HIV载体浓度达56672．93pg/ml,荧光显微镜下Hela细胞胞浆内充满深浅不一的绿荧光,大鼠脊髓神经干细胞球发强烈的绿荧光。结论此实验方法操作简单、快捷,具有高效性及安全性。HIV载体将是基因治疗的理想工具。

Objective Introducing experiment methods of constructing lentivirus vector deriving from human immunodeficieney virus-1 （ HIV-1 ） and transferring it into rat spinal cord neural stem cells. Methods GFP-HIV vector was assembled in 293T line with 4 plasmids containing green fluoreseence protein （GFP） , the high concentration and intense infectivity of GFP-HIV vector were checked with Elisa Kit and Hela line respectively, and then transferred to rat spinal cord neural stem cells. Results The concentration of GFP-H1V was 56672.93 pg/ml, green fluorescence were seen in Hela cells and neural stem cell spheres. Conclusions The experiment methods are convenient, which have intense infectivity and safety. HIV vector is an excellent tool for experimental gene transfer and promising candidate for gene therapy.