Triton X-100对脂质体介导的BMP-2基因转染大鼠BMSCs的作用

发布时间：2018-03-22 23:28

本文选题：BMSCs　切入点：Triton　出处：《中国修复重建外科杂志》2015年01期 　论文类型：期刊论文

【摘要】：目的探讨Triton X-100对脂质体介导的BMP-2基因转染大鼠BMSCs的作用。方法 8周龄Wistar大鼠1只,雄性,体质量120 g。取Wistar大鼠股骨、胫骨骨髓,采用贴壁培养法分离培养BMSCs并鉴定。取第3代BMSCs,采用MTT法筛选Triton X-100适宜浓度。实验分成3组,实验组:BMP-2+脂质体+0.010%Triton X-100+BMSCs,常规转染组:BMP-2+脂质体+BMSCs,空白对照组:BMSCs+血清培养基。转染后48 h,倒置荧光显微镜观察各组绿色荧光蛋白表达情况;转染后72 h,采用实时荧光定量PCR检测BMP-2 m RNA表达。结果 0.010%Triton X-100既能使BMSCs保持一定的活力,又可达到一定的作用效果,故确定其为适宜浓度。转染后48 h,倒置荧光显微镜下见实验组和常规转染组均有绿色荧光蛋白表达,空白对照组无绿色荧光蛋白表达。转染后72 h,实验组BMP-2 m RNA相对表达量为5.94±0.12,常规转染组为4.99±0.08,差异有统计学意义(t=360.28,P=0.02)。实验组转染效率较常规转染组平均提高了19%。结论 0.010%Triton X-100可以促进脂质体介导的BMP-2基因转染大鼠BMSCs,并提高其转染效率。
[Abstract]:Objective to investigate the effect of Triton X-100 on liposome-mediated BMP-2 gene transfection into rat BMSCs. Methods the bone marrow of femur and tibia of Wistar rats was extracted from femur and tibia bone marrow of 8 week-old Wistar rats (male, 120g). BMSCs was isolated and identified by adherent culture. The optimal concentration of Triton X-100 was screened by MTT method and the third generation BMSCs were selected. The experiment was divided into 3 groups. 0.010%Triton X-100 BMSCs, 0.010%Triton X-100 liposome in the experimental group, BMSCs in the conventional transfection group and in the blank control group, respectively. 48 hours after transfection, the expression of green fluorescent protein (GFP) in each group was observed by inverted fluorescence microscope. The expression of BMP-2 m RNA was detected by real-time fluorescence quantitative PCR at 72 h after transfection. Results 0.010%Triton X-100 could not only maintain the activity of BMSCs, but also achieve a certain effect. After 48 hours of transfection, green fluorescent protein expression was found in both experimental group and conventional transfection group under inverted fluorescence microscope. The relative expression of BMP-2 m RNA was 5.94 卤0.12 in the experimental group and 4.99 卤0.08 in the conventional transfection group at 72 h after transfection, the difference was statistically significant. The transfection efficiency of the experimental group was higher than that of the conventional transfection group. 0.010%Triton X-100 could promote liposome-mediated BMP-2 gene transfection into BMSCs and improve its transfection efficiency.
【作者单位】：泸州医学院附属医院口腔颌面外科;
【基金】：四川省科技厅应用基础研究资助项目(2008jy0014)~~
【分类号】：R782

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