Optimization of Condition for Transfection of Jurkat Cells with Lipofectamine~ LTX

Objective To optimize the condition for transfection of Jurkat cells with Lipofectamine LTX.Methods Jurkat cells were transfected with Lipofectamine LTX using plasmids pIRES2-EGFP and pIRES2-EGFP-STAT6,both carrying green fluorescent protein(GFP) gene,as vectors,based on which the cell growth curve was plotted,the effect of freshness of medium on cell growth as well as effects of time for culture,ratio of Lipofectamine LTX to plasmid DNA,density of cells inoculated and volume of medium on transfection efficacy were evaluated,and the transient transfection efficacies were compared.Results The growth rate of Jurkat cells was closely related to the cell density.The change of medium 1 d before transfection ensured the normal growth state of cells.However,the freshness of medium showed no significant effect on cell growth.When the ratio of Lipofectamine LTX volume to plasmid mass was 2.75 μl ∶ 500 ng,the transfection efficacy was satisfactory.Properly increased density of Jurkat cells increased the utilization rate of transfection reagent.Forty-eight hours after transfection,the transfection efficacy of cells inoculated in 300 μl of medium onto each well of culture plate was(1.4 ± 0.2) times of that in 600 μl of medium,which showed significant difference(P 0.05).The transfection efficacies at various ratios of Lipofectamine LTX volume to plasmid mass showed no significant difference 24 h(P 0.05) while showed significant difference 48 h(P 0.05) after transfection.The transfection efficacy of plasmid pIRES2-EGFP was significantly superior to that of plasmid pIRES2-EGFP-STAT6(P 0.05).Conclusion The condition for transfection of Jurkat cells with Lipofectamine LTX was optimized,which provided a reference for the transfection of suspension cells.