超声作用对绵羊胎儿成纤维细胞转染效率的影响

为研究超声波对经脂质体介导的绵羊胎儿成纤维(sheep fetal fibroblasts,SFF)细胞转染效率的影响,用不同频率的超声波在不同时间内辐射经脂质体介导的SFF细胞,转染48h后收集细胞,用RT-PCR检测目的基因,台盼蓝染色检测细胞活性。转染后用G418挑选转基因单克隆细胞,计算转染率。结果显示,对细胞、质粒和脂质体的混合液共同超声辐射(时间60s,超声频率为2.5MHz)后转染,转染率要明显高于未超声辐射的细胞和先转染后超声辐射的细胞,并且细胞的存活率较高。结果表明,超声辐射不仅能改变细胞质膜的通透性,还能够提高脂质体介导的基因转染效率,增强基因介导的特异性和靶向性。

Effect of ultrasound on transfection efficiency of sheep fetal fibroblasts

This study was designed to investigate the influence of ultrasound on the lipofectamine conducted sheep foetus fibroblasts(SFF) cell transfection efficiency.The lipofectamine conducted SFF cells were subjected to ultrasound with different time and frequency,collected 48 hours after transfection.The target gene was detected by PCR and RT-PCR and the cyto-activity was determined by trypan blue stain.G418 was used for culling the transgenes,monoclonal cells and counting transfection efficiency.In result,the cells and their plasmids and lipofectamine exposed(Exposure time is 60 s under 2.5 MHz) together would like to lead to a higher transfection efficiency than the cells without exposure cells.In conclusion,the ultrasound exposure can not only improve the cell’s permeability,but also the lipofectamine-conducted gene transfection efficiency,and enhance the specificity and targeting effect of gene-conducted transfection.