Experimental Study on a New Method for Retrovirus Titer Determination

[Objective] To investigate a convenient and effective in vitro cell culture method to acquire high retrovirus titer.[Methods] The retrovirus vector PLXIN were transfected to PA317 packaging cells by liposomal transfection reagent,and were selected by G418 and expanded culturing.Then the supernanant was collected and the Virus titers were detected.To compare the retrovirus quantity generated by different methods can establish better experimental basis for improving the titer and further infecting target cell.[Results] Compared with the normal metuods(37℃,5% CO2),the new improved method(low temperature32℃,CO2 non-existence and by using frozen-melting method)can enhanced the retrovirus titer at lease 10 times.PTs,PTas and PLXIn group retrovirus vector are enhanced to 1.3×105CFU/mL,1.2×105CFU/mL and 1.5×105 CFU/mL respectively.[Conclusion] New method for retrovirus titer determination is convenient and effective,by which higher virus titers can be acquired.