Construction of EGFP Expressing HepG2 Cell Line Using Electroporation

The human hepatoma HepG2cell line has been extensively used as an experimental model in toxicological and pharmacological research. HepG2 cells, similar to primary hepatocytes, are difficult to transfect with traditional liposome- based methods. Electroporation is a physical method for cell transfection, where application of high voltage electric pulses is used to transiently permeabilize cell membranes allowing facilitated entry of plasmid DNA into the cells. An optimized electroporation protocol was developed for transfection of HepG2 cells stably expressing green fluorescent protein. Electric pulses of 600 V/cm yielded the highest transfection efficiency and stably transfected clones were selected after 14 days incubation in selection medium. High correlation between emitted fluorescence and different cell densities were demonstrated. Such stably transfected cells HepG2-EGFP can be used for the development of rapid and simple cytotoxicity assay, that can be easily amenable for automation in highthroughput screening setups.