Optimization of Electroporation Parameters for Cells Cultured in Suspension

Objective To optimize the electroporation parameters in leukemia cell lines cultured in suspension using GFP (green fluorescent protein) as a reporter gene. Methods The GFP plasmid was transferred into leukemia cell lines K562 and NB4 by electroporation using differently experimental conditions such as the voltage, the electric capacity,the state of cell vitality, and the operation tempe rature. The transfection efficiencies were evaluated by flow cytometry(FCM) and fluorescent microscopy. Results The highest electroporation transfection rate was achieved under the condition of voltage 250 V or 350 V,electric capacity 950 μF, 40.6% and 32.9% for these two cell lines respectively. The transfection efficiencies were closely related to the vigorous state of the cells. The operation temperature and the concentration of serum in buffer have no effect on the transfection efficiencies.Conclusion Electroporation is a highly efficient gene transfection method. Optimized electroporation parameters can improve the transfection efficiency.