Effect of hypotonic stress on retroviral transduction.
Short half-life has long been known as a main barrier for retroviral gene delivery due to quick degradation that seriously limited application of retrovirus-mediated methodology in the clinical use. To circumvent this challenge, many physical and chemical approaches have been developed to maximize contact opportunity of retroviruses and cells before viral vectors decay. However, most of methods are not easy to be followed due to complicated equipment settings and/or long procedures. In this study, we introduced an easy, cost-effective, efficient, and scalable strategy to enhance retroviral transduction by hypo-osmotic stress. It has been demonstrated that under hypotonic exposure, cell membrane is permeabilized to allow numerous exterior molecules accessing to cytoplasm through an intensive endocytosis, yielding high efficiency of cellular uptake. We hypothesized this hypotonic stress-induced internalization may provide a unique opportunity of cell entry for retroviruses without the need of receptor binding, and thus overcome the insufficient transduction rate due to loss of envelope protein. Indeed, our results showed that with assistance of hypotonic stress, retroviral transduction rates dramatically increased about 5.6- and 17.7-fold using fresh and decayed retroviruses, respectively, in comparison with corresponding groups without hypotonic stress. In summary, hypotonic stress was shown as a promising tool for enhancement of retroviral transduction efficiency without limitation of short half-life.
Lee
Yu-Hsiang
YH
Department of Chemical Engineering, University of Southern California, Los Angeles, CA, USA.
Peng
Ching-An
CA
D000818:Animals; D002463:Cell Membrane Permeability; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D007038:Hypotonic Solutions / Q000494:pharmacology; D051379:Mice; D041681:NIH 3T3 Cells; D009997:Osmotic Pressure; D012190:Retroviridae; D014161:Transduction, Genetic / Q000379:methods*
D007038:Hypotonic Solutions