Effects of Bcl-xl/Lipofectamine ~(TM)2000 ratio on the transfection efficiency in human keratocyte

Objective To evaluate the best ratio of Bcl-xl/Lipofectamine TM2000 for human keratocyte transfection.Methods Different concentrations of Bcl-xl and Lipofectamine TM2000 was mixed to make Bcl-xl/Lipofectamine TM2000 complex,with the final Bcl-xl/Lipofectamine TM2000 ratios of 1∶2,1∶4,1∶8 and 1∶10 respectively.All the Bcl-xl/Lipofectamine TM2000 complex were incubated with cultured human keratocyte for 24 hours before transfection.The transfection efficiency was tested with RT-PCR,immunohistochemistry,flow cytometry methods on the 3rd day of transfection.The data was analyzed with SPSS software.Results The transfection efficiency of Bcl-xl gene was various dependent on the Bcl-xl/Lipofectamine TM2000 ratio.When Bcl-xl/Lipofectamine TM2000 ratio was 1∶8,the highest transduction efficiency appeared as high as (45.6±2.0)% (immunohistochemistry),(48.3±3.0)%(flow cytometry),significantly higher than other groups(P0.05).Conclusion Under appropriate conditions,cationic liposome can facilitate the transfection of Bcl-xl gene into keratocyte.The best ratio of Bcl-xl/Lipofectamine TM2000 was 1∶8.