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Optimization of transfection conditions of CTGF shRNA plasmids into pterygium fibroblasts

Luo Liqing,Wu Ping,Liu Yanyan,He Huijuan,Wang Sijie,Wu Weiquan (Research Center of Medical Science, Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)  
Objective To investigate the optimal ratio of plasmids to Lipofectamin 2000 after CTGF shRNA plasmids were transfected to pterygium fibroblasts.Methods The primary pterygium fibroblasts were cultured using the method of attachment tissues,and identified with immunohistochemical SP method.Cells of the second or third generation were transfected by using different ratios of CTGF shRNA plasmids to Lipofectamin 2000.After 24 h,the cell transfection efficiency was analyzed by flow cytometry.Then the optimal ratio of plasmids to Lipofectamin 2000 was determined according to the highest transfection efficiency.Results The optimal ratio of plasmids to Lipofectamin 2000 was 1∶2.5,and the transfection efficiency was (56.7±1.5)%.Conclusion This experiment might lay the foundation for performing efficient in vitro transfection into cultured cells and further research on RNA interfere of CTGF gene.
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