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《Chinese Journal of Reparative and Reconstructive Surgery》 2015-01
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EFFECT OF TRITON X-100 ON LIPOSOME MEDIATED BONE MORPHOGENETIC PROTEIN 2 BY TRANSFECTION OF RAT BONE MARROW MESENCHYMAL STEM CELLS

XIA Delin;HUANG Mingke;FU Guangxing;MA Zheng;WU Shuangjiang;ZHOU Hangyu;Department of Oral and Maxillofacial Surgery,the Affiliated Hospital of Luzhou Medical College;  
Objective To study the effect of Triton X-100 promoting liposome-mediated bone morphogenetic protein 2(BMP-2) gene transfection of rat bone marrow mesenchymal stem cells(BMSCs). Methods BMSCs were separated and cultured from the femur and tibia of healthy Wistar rats(8-week-old,male). The 3rd passage BMSCs identified by detecting the surface antigen were used to transfect. The optimum concentration of Triton X-100 for liposome mediated gene transfection was determined with ELISA meter by the way of MTT. In optimum concentration of Triton X-100,liposome mediated BMP-2 gene was transfected to BMSCs. The experiment was divided into 3 groups according to types of trasfection agents: BMSCs were transfected with Triton X-100+liposome+BMP-2(experimental group),with liposome+ BMP-2(conventional transfection group),and untransfected BMSCs served as blank control group. After 48 hours of transfecting,the green fluorescent protein(GFP) in cells was detected through inverted fluorescence microscope. After 72 hours of transfection,real-time fluorescence quantitative PCR was applied to measure the m RNA expression of BMP-2. Results 0.01% Triton X-100 was determined to be the optimum concentration for not only making the BMSCs maintain vitality,but also achieving a certain effect on BMSCs. After trasfecting for 48 hours,GFP was observed through inverted fluorescence microscope in the experimental group and conventional transfection group,but was not observed in the blank control group. After trasfecting for 72 hours,the relative BMP-2 m RNA expression level was 5.94±0.12 in the experimental group,and was 4.99±0.08 in the conventional transfection group,showing significant difference(t=360.28,P=0.02). The transfection efficiency was increased by 19% in the experimentalgroup. Conclusion 0.010%Triton X-100 can promote the liposome mediated BMP-2 gene transfection of rat BMSCs,and can improve the transfection efficiency.
【Fund】: 四川省科技厅应用基础研究资助项目(2008jy0014)~~
【CateGory Index】: R782
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