优化人单核细胞型淋巴瘤THP1细胞的电转条件

Optimization of electroporation conditions of human monocytes THP1 cells

目的优化THP1细胞的电转条件。方法首先检测不同培养基在不同体积下的电阻，其次调整不同的电压和电容的组合，最后测试质粒质量对转染效率的影响。结果THP1细胞完全培养基、无血清培养基和OPTI—MEMI在同一体积下的电阻一致，同一种培养基在不同体积下体积越大电阻越小；适宜的电压和电容分别为300V和150μF；质粒质量对电转的影响表现为20肛g质粒的转染率明显高于15μg、25μg和30μg。结论本实验从培养基、电压、电容以及质粒质量这三个方面成功地优化了THP1细胞的电转条件。

Objective Optimization the electroporation conditions of THP1 cells. Methods First,we test the resistances of dif- ferent medium in different volumes. Second,we adjust the different combinations of voltage and capacitance. At last,we detected the effect of different quality of plasmid in electroporation. Results The resistances of complete medium,FBS-free medium and OPTI- MEMI were similar in the same volume; However for the same medium,the smaller volume,the greater the resistance. Voltage and capacitance were 300V and 150/2F. The effect of plasmid quality for electroporation efficiency was that 20μg plasmid transfection rate was higher than 15μg,25μg and 30μg. Conclusion In present experiment,the electroporation conditions of THP1 cells were successfully optimization by the choice of medium,voltage,capacitance and plasmid quantity.