| Objective Superfect is a sort of novel nanoscopic vector, which transfection efficiency is related to many factors such as the dose of DNA,N/P ratio( N: number of amidocyanogens on the surface of vector.P: number of phosphate groups of DNA ),cell species.cell density and degree of confluency, time of Superfect- DNA complex co-incubating with cells,and so on. We design the experimentation to do primary study of transfection efficiency , cytotoxicity and optimized transfection condition of Superfect? then discuss the value of its application in the field of gene therapy.Methods The murine melanoma cell line, B16F0, with 50% cell confluency rate is the target cell and the enhanced green fluorescent protein (EGFP) expressing plasmid, pEGFP-Cl, is the input gene. Various transfection protocols are carried in experiment groups,i.e.two DNA dose(0.25ug,0.50ug),three N/P ratio(2:l,5:1,10:l) and two complex co-incubating with cell(3h,6h).The control group is composed of naked gene or vector transfection group and normal group.The efficiency of transfection is examined by Flow CytoMeter and the cytotoxicity is evaluated with MTTassay.Results There is significant difference among experiment groups(P<0.05). Superfect?shows some cytotoxicity in the groups treated with high N/P ratio and long time of co-incubation. Superfect?has higher transfection efficiency and lower cytotoxicity in the group of 0.5ug plasmid,5:1 N/P ratio and 3 hours' incubating with cells(P<0.05). It also suggests the increasing trend of protein expression after transfection in 6 day's observation.Conclusion Superfect is the vector with high transfer efficiency and low cytotoxicity.Its optimized transfection condition and influence on cell survival is mainly related to N/P ratio and time of co-incubation.Continuous expression of green fluorescent protein suggests that the vector is possible to mediate stable transfection of DNA. Superfect possesses of well application foreground in the field of gene therapy.
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