Detection of TIMP-1 Targeted siRNA Transfection Efficiency in Hepatic Stellate Cell

To increase the transfection efficiency of TIMP-1 pGCsi-U6/siRNA expression constructs transfection in HSC-T6 cell line.HSC-T6 cell lines were transfected with TIMP-1 pGCsi-U6/shRNA plasmid DNA.According to the different ratios of plasmid DNA/Lipofectamine 2000(Life technologies),the transfection protocols were divided into 5 groups:naked DNA,2 μg/4 μL、2 μg/6 μL、2 μg/8 μL,2 μg/10 μL;48 hours later,transfection efficiencies of different protocols were evaluated by Laser Scanning Confocal Microscope observation and flow cytometer counting.The best transfection efficiency could be got when the ratio of plasmid DNA/Lipofectamine 2000 is 2 μg/8 μL compared with the rest groups.It had significant difference,which showed that transfer efficiency was correlated with the ratio of plasmid to lipofectamine.The efficiency of transferring TIMP-1 pGCsi-U6/shRNA into HSC-T6 can achieve to 37% with proper ratio of plasmid to lipofactamine.TIMP-1 pGCsi-U6/shRNA are ideal transient expression vectors for HSC-T6 gene transference.