Abstract:

BACKGROUND: Recently, cationic liposome becomes the commonly used vehicles for gene transfer. However, few study reports transfection of mIMCD-3 cell lines mediated by liposome.
OBJECTIVE: To approach the feasibility of transfection of mIMCD-3 mediated by liposome and to obtain the optimized transfection condition for mIMCD-3.
METHODS: Using green fluorescent protein as reporter gene, the plasmid pIRES2-EGFP was transfected into mIMCD-3 cells with LipofectamineTM 2000. To set different groups according to cell density, DNA amount, ratio of DNA and liposome, plasmid extraction methods and serum, and to observe the influences on transfection efficiency by these different factors.
RESULTS AND CONCLUSION: The highest transfection efficiency was achieved with the following optimized conditions:  2×10⁵/ L cells, 1 g DNA, 1:3 ratio of DNA and liposome. Extracting plasmid by E.Z.N.A. Endo-Free Plasmid Mini Kit produced by American OMEGA company obtained higher transfection efficiency. Compared with serum medium, cells culutred with serum-free medium obtained higher transfection efficiency before transfection (P < 0.01), but it had no effect at 6 hours after transfection. Transfection of mIMCD-3 mediated by liposome is feasible, and the optimized transfection condition for mIMCD-3 is obtained. These results may be used as the reference for other studies．