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Centrifugation Increases the Gene Transfection Efficiency of HaCaT Cells
Xu Yunfei1, Meng Shasha1, Zhou Weihui1*, Li Hongmei2*
1Pediatric Research Institute, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing 400014, China; 2Department of Dermatology, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China
Abstract: HaCaT cells are spontaneous human immortalized epidermal cell lines that are commonly used in the study of skin function and disease but the efficiency of conventional liposome transfection of the cells is extremely low. This study is aimed to investigate whether centrifugation can increase the efficiency of liposomal transfection of HaCaT cells. Lipofectamine® 2000 was used as a liposomal transfection reagent， and pEGFP-C1 was used as a plasmid， and HaCaT cells were transfected by conventional method and centrifuged method respectively. Transfection efficiency was observed by inverted fluorescence microscope， quantitatively determined by flow cytometry. Cell proliferation activity was measured to determine whether centrifugation affect cell viability. The luciferase reporter assay and Western blot were carried out to detect the effect of centrifugation on the expression of the target gene. The results showed that the transfection efficiency of centrifugation treatment was higher than that of the conventional treatment. The difference was statistically significant. Luciferase reporter gene assay and Western blot experiment further confirmed that the centrifugation after transfection could promote the expression of the target gene， which had high practicability. In summary， the transfection efficiency of HaCaT cells can be increased by centrifugation after conventional transfection procedures.
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