Effective parametric optimization to improve transfection efficiency of CHO cells via electroporation

Abstract

<p>The establishment of effective gene transfer method leads to the improvement in gene therapy protocols. The electroporation technique has been positively accepted for in vitro and in vivo studies, but its poor transfection efficiency is still a challenging task. Hence, the attention of researchers is to increase the transfection efficiency while maintaining cell viability. The electrotransfection conditions for each cell type should be determined to achieve the maximum plasmid transfer while preventing the undesirable cell deaths. In this research, the optimal values of electrotransfection parameters for in vitro transfection of CHO cells are determined. Some of these varying parameters are the number of applied pulses and plasmid DNA concentration at which the electroporation was carried out. GFP coding plasmid concentrations of 10, 100, 200, 300, 400 and 600 μg/ml were chosen. Different tendencies of transfection efficiency were observed for different HV pulses (1, 2 and 3 HV) with higher (400 and 600 μg/ml) plasmid concentration. For all the three pulse combinations the cell viability was continuously decreased with increase in plasmid concentration from 10, 100, 200, 300, 400 and 600 μg/ml. The increase in plasmid concentration from 10, 100, 200, 300, 400 and 600 μg/ml increased the corresponding transfection to 6.78, 33.99, 49.34, 65.52, 71.61 and 59.36 % in the case of 3 HV pulses. The optimal value of plasmid concentration for electrotransfection was found from the presented dynamics of cell viability decline and transfection efficiency rise when increasing the plasmid concentration.</p>