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The experimental study of genetic engineering human neural stem cells mediated by lentivirus to express multigene

机译:慢病毒介导人神经干细胞表达多基因的基因工程实验研究

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Objective: To explore the feasibility to construct genetic engineering human neural stem cells ( hNSCs) mediated by lentivirus to express multigene in order to provide a graft source for further studies of spinal cord injury (SCI). Methods: Human neural stem cells from the brain cortex of human abortus were isolated and cultured, then gene was modified by lentivirus to express both green fluorescence protein ( GFP) and rat neurotrophin-3 (NT-3); the transgenic expression was detected by the methods of fluorescence microscope, dorsal root ganglion of fetal rats and slot blot. Results: Genetic engineering hNSCs were successfully constructed. All of the genetic engineering hNSCs which expressed bright green fluorescence were observed under the fluorescence microscope. The conditioned medium of transgenic hNSCs could induce neurite flourishing outgrowth from dorsal root ganglion (DRG). The genetic engineering hNSCs expressed high level NT-3 which could be detected by using slot blot. Conclusions : Genetic engineering hNSCs mediated by lentivirus can be constructed to express multigene successfully.
机译:目的:探讨构建慢病毒介导的基因工程人神经干细胞(hNSCs)以表达多基因的可行性,为进一步研究脊髓损伤(SCI)提供移植来源。方法:分离并培养人流产大脑皮层的人神经干细胞,然后用慢病毒修饰基因使其表达绿色荧光蛋白(GFP)和大鼠神经营养蛋白3(NT-3)。用荧光显微镜,胎鼠背根神经节和缝隙印迹法检测转基因表达。结果:成功构建了基因工程hNSC。在荧光显微镜下观察到所有表达亮绿色荧光的基因工程hNSC。转基因hNSCs的条件培养基可以诱导背根神经节(DRG)的神经突生长。基因工程hNSCs表达高水平的NT-3,可以通过缝隙印迹检测到。结论:慢病毒介导的基因工程hNSCs可成功表达多基因。

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