The Optimization of Transfection Conditions for Rat BHK Cells and Chinese Hamster BRL Cells

Objective:To optimize the Lipofectamine 2000 transfection conditions for rat BHK cells and Chinese hamster BRL cells using β-Gal vectors.Methods: Lipofectamine 2000 were mixed in varied ratios with plasmid DNA to transfect the rat BHK cells and Chinese hamster BRL cells on a 96-well plate.Successfully transfected cells were stained blue due to expression of β-Gal degradable substrates(X-gal),enabling estimation of the transfection efficacy by counting the blue cells under microscope.ANOVA was performed for statistical test.Results: The maximal efficacy was(43.2±9.3)% in BHK cells when β-Gal vector and Lipofectamine 2000 were mixed in a ratio of 0.27∶0.8(μg∶μL),and was(5.7±1.3)% in BRL cells when β-Gal vector and Lipofectamine 2000 were mixed in a ratio of 0.10∶0.3(μg∶μL).Conclusion: Lipofectamine 2000 appears to be useful for the transfection of mammal cells;however,the transfection conditions should be optimized for individual cell types.