【摘要】 目的以原代培养的SD大鼠神经干细胞为研究对象,优化电穿孔转染条件。方法通过设置不同的电压、脉冲时间、电压脉冲时间组合等电转染条件,将表达绿色荧光蛋白(GFP)的靶向大鼠β-catenin基因的siRNA质粒导入SD大鼠神经干细胞,48 h后观察并比较神经干细胞的转染效率,确定最佳电转染条件。结果 300 V电压、60μs脉冲时间或300 V电压、80μs脉冲时间的转染条件下,SD大鼠神经干细胞可获得较好的转染效率,分别为(62.21±11.56)%和(53.34±9.0)%。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SD大鼠神经干细胞的电转染效率更多还原

【Abstract】 【Objective】 To optimize the electroporation parameters in primary cultured SD rats neural stem cells.【Methods】 Green fluorescent protein(GFP) expression plasmid siRNA was transferred into neural stem cells using different electroporation Voltages,impulse times,times et al.Expression of GFP was observed after forty-eighty 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 300 V,pulse time 60 μs or voltage 300 V,pulse time 80 μs for the primary cultured SD rats neural stem cells.The transfection rate was(62.21±11.56) % and(53.34±9.00) % respectively.【Conclusion】 Electroporation is a highly efficient gene transfection method.Optimized electroporation parameters can improve the electroporation efficiency of the SD rats neural stem cells.更多还原