小鼠大脑皮质神经元电穿孔转染条件的优化

Optimization of electroporation parameters for cerebral cortical neurons of mouse

目的:以原代培养的小鼠大脑皮质神经元为研究工具,优化电穿孔转染条件。方法:通过设置不同的电压和脉冲时间组合等电转染条件,将表达绿色荧光蛋白(GFP)的shNC质粒导入小鼠大脑皮质神经元,24 h后观察并比较神经元的转染效率,确定最佳电转染条件。结果:200 v电压,25 ms脉冲时间或250 v电压,15 ms脉冲时间的转染条件下,小鼠大脑皮质神经元可获得较好的转染效率,分别为(31.15±1.89)%和(29.10±1.93)%。结论:电穿孔转染是一种高效的基因转染方法,通过条件的优化,可以提高小鼠大脑皮质神经元的电转染效率。

Objective： To optimize the electroporation parameters in primary cultured cerebral cortical neurons of mouse.Methods： Green fluorescent protein（GFP） expression plasmid shNC was transferred into cortical neurons using different electroporation voltages and impulse times.Expression of GFP was observed after twenty-four hours of transfection and the electroporation efficiencies were analyzed to determine the optimal electroporation conditions.Results： The highest electroporation transfection rate was achieved under the conditions of voltage 200 v,pulse time 25 ms or voltage 250 v,pulse time 15 ms for the mouse cortical neurons.The transfection rate was（31.15±1.89）% and（29.10±1.93）% respectively.Conclusion： Electroporation is a highly efficient gene transfection method.Optimized electroporation parameters can improve the electroporation efficiency of the mouse cortical neurons.