The Dependence of electrotransfection efficiency on the duration of cell growth passage

Abstract

ONLINE ISSN: 2335-8718In clinical applications, such as DNA vaccination and gene therapy, gene electrotransfer is used as one of the most promising and efficient technique. The proper establishment of the gene transfer method ensures the improvement of gene therapy protocols. Electroporation has been increasingly employed in in vitro and in vivo protocols, and much attention has been given to increasing its transfection potential. Electroporation is a relatively safe and simple technique to deliver nucleic acids to the cell that acts by rendering cells transiently permeable using short periods of high voltage. To achieve the maximal introduction of plasmid DNA into cells and, at the same time, to prevent undesirable cell deaths, electro transfection conditions should be determined for every single cell type individually. In the present study, we determined the duration of cell growth passage for in vitro transfection of CHO cells. Time of 24 h and 48 h cell passage before the experiment was chosen. Electrotransfection efficiency with all plasmid concentrations significantly differed when comparing 24 h and 48 h passage time. Transfection efficiency from 47.28±0.41% after 24 h passage time fell to 19.87±1.02% after 48 h of cell passage time using same electric field parameters. But there was very less viability change when comparing cell passage time of 24h and 48 h. The cell passage time, if optimized, may generate a reproducibly high proportion of transfected cells among the different cell typesBiologijos katedraGamtos mokslų fakultetasVytauto Didžiojo universiteta