Optimization of transfection conditions for tranfecting siRNA to EOMA cells mediated by cationic liposome

Objective To optimize the transfection parameters for transfecting siRNA(small interfering RNA) to EOMA(endotheliumof mouse angioma) cells mediated by LipofectamineTM2000.Methods 1.0μl and 1.5μl of LipofectamineTM2000 diluted with50μl DMEMwere mixed with different concentrations of fluorescin-labeledsiRNAtoforma series of mixturestotransfect EOMAcells.24 h after transfection,cell count was made under fluorescent microscope to determine the percentageof the cells transfected under each condition.The cell viability was calculated by using MTTassay at the same time.Results No differences werefoundintransfection efficiency and cell viability whether using1.0μl or 1.5μl LipofectamineTM2000,if theamount of siRNAwasfixed.WhensiRNAwas 50 pmol,the percentage of positive cells was as high as 86 %.Thetransfection ef-ficiency could not befurtherincreased by elevatingthe concentration of siRNA,and 60 pmol siRNAwould bring about higher cy-totoxicity.Conclution The transfection efficiency and cell viability are dependent onthe correct LipofectamineTM2000-siR-NAratio.