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目的观察阳离子脂质体法转染体外培养神经干细胞的转染效率和外源基因的表达。方法从1d龄新生鼠大脑皮层组织培养神经干细胞,带有报告基因GFP的穿梭质粒pAdTractCMV经Lipofectamine介导转染神经干细胞后观察GFP表达,用流式细胞仪测定转染率。并观察阳离子脂质体对神经干细胞的毒性作用。结果荧光显微镜观察到被转染的神经干细胞长期表达绿色荧光蛋白。流式细胞仪结果显示转染率最高可达到39.99%。转染时,阳离子脂质体浓度超过24ml/L时表现出细胞毒性。结论阳离子脂质体lipofectamine介导转染神经干细胞效率较高,外源基因表达时间长。
Objective To observe the transfection efficiency and the expression of exogenous gene of neural stem cells transfected by cationic liposomes in vitro. Methods Neural stem cells (NSCs) were cultured from cerebral cortex of 1-day old neonatal rats. The shuttle plasmid pAdTractCMV with reporter gene GFP was transfected into neural stem cells by Lipofectamine to observe the expression of GFP. The transfection efficiency was determined by flow cytometry. And observe the cationic liposomes on the toxic effects of neural stem cells. Results Fluorescence microscopy showed that transfected NSCs express green fluorescent protein for a long time. Flow cytometry results show that the highest transfection rate can reach 39.99%. When transfected, the cationic liposomes showed cytotoxicity when the concentration exceeded 24 ml / L. Conclusion The lipofectamine-mediated transfection of neural stem cells is more efficient and the expression of foreign genes is longer.