Enhanced retroviral transduction of 293 cells cultured on liquid-liquid interfaces

In clinical research, retrovirus-mediated gene therapy is one of the most c ommonly used methods to deliver and express the gene of interest due its ab ility to allow for stable gene integration into the chromosomes of target c ells. To elevate the efficiency of viral transduction, several restrictions , such as low virus-cell encounters and the necessity for cell division, mu st be improved. In this study, we focused on the possibility of acceleratin g cell division and the ensuing increment of viral transduction on flexible substrata. Perfluorocarbon FC-40 was harnessed to form a liquid-liquid int erface with culture medium. Enhanced green fluorescence protein (EGFP) was employed as the marker gene to quickly illustrate the percentage of viral i nfection. The results indicate that the gene transfer efficiency to 293 cel ls cultured on protein-precoated liquid-liquid interfaces was higher than i n cells cultured on rigid polystyrene surfaces. This increased transduction rate on the liquid-liquid interface is consistent with the acceleration of division of 293 cells on a flexible interface, which exhibited less adhesi veness. The effect of cell-cell contact inhibition on the rate of gene tran sduction is also addressed in this study. (C) 2001 John Wiley & Sons, Inc.