Lipofectamine介导转染神经干细胞的研究

Neural stem cell transfection through lipofectamine

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 investigate the expression of extrinsic gene in neural stem cell (NSC) and the cytotoxicity of lipofectamine after transfection through lipofectamine.Methods The neural stem cells were culture and transfected with pAd-GDNF plasmid through lipofectamine.The expression of green fluorescent protein (GFP) was detected by fluorescent microscopy and flow cytometer.Results The transfected neural stem cells expressed GFP for a long-term.Flow cytometry revealed that the highest transfection rate was up to 39.90%.The lipofectamine became toxic to NSCs when its concentration was over 24 ml/L during the process of transfection.Conclusion The expression of extrinsic gene in NSC could be a long-term mode.Transfection of neural stem cell mediated by lipofectamine is effective and the extrinsic gene has a long-term expression.