Enhanced retroviral transduction of 293 cells cultured on liquid-liquid interfaces.
In clinical research, retrovirus-mediated gene therapy is one of the most commonly used methods to deliver and express the gene of interest due its ability to allow for stable gene integration into the chromosomes of target cells. To elevate the efficiency of viral transduction, several restrictions, such as low virus-cell encounters and the necessity for cell division, must be improved. In this study, we focused on the possibility of accelerating cell division and the ensuing increment of viral transduction on flexible substrata. Perfluorocarbon FC-40 was harnessed to form a liquid-liquid interface with culture medium. Enhanced green fluorescence protein (EGFP) was employed as the marker gene to quickly illustrate the percentage of viral infection. The results indicate that the gene transfer efficiency to 293 cells cultured on protein-precoated liquid-liquid interfaces was higher than in 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 adhesiveness. The effect of cell-cell contact inhibition on the rate of gene transduction is also addressed in this study.
Kwon
Y J
YJ
Department of Chemical Engineering, University of Southern California, 925 West 37th Street, Los Angeles, California 90089-1211, USA.
Yu
H
H
Peng
C A
CA
D002448:Cell Adhesion; D002455:Cell Division; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D005466:Fluorocarbons / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics; D008562:Membrane Glycoproteins; D008856:Microscopy, Fluorescence; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000378:metabolism; D011137:Polystyrenes / Q000378:metabolism; D013499:Surface Properties; D014161:Transduction, Genetic / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D005466:Fluorocarbons; C028391:G protein, vesicular stomatitis virus; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D011137:Polystyrenes; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins; D005947:Glucose