【摘要】 目的探讨超声微泡介导pGPU6/Neo质粒转染神经干细胞(NSCs)的效率及可行性。方法以一定能量的超声介导质粒转染大鼠NSCs,分为:空白组、质粒组、质粒+微泡组、质粒+超声组、质粒+微泡+超声组,并按不同转染条件分为亚组。转染48 h后荧光显微镜观察转染效率,台盼蓝染色法检测细胞活性。结果最优参数为声强1.0 w/cm~2,辐照时间30s时,超声微泡介导质粒转染率最高,且对细胞活性无明显影响。超声微泡介导质粒对NSCs的转染效率,明显高于其他实验组(P<0.05)。结论在适当超声强度和辐照时间条件下,超声微泡能够安全、有效地介导基因转染NSCs。更多还原

【Abstract】 Objective To investigate the efficiency and feasibility of pGPU6/Neo plasmid transfected into neural stem cells(NSCs)by ultrasound-mediated microbubble destruction.Methods The plasmid was used to transfect into NSCs by ultrasound-mediated microbubble destruction.The NSCs were divided into 5 groups:blank control,plasmid, plasmid-microbubble,plasmid-ultrasound and plasmid-microbubble-ultrasound.The last group was further divided into subgroups according to different conditions.After 48 hours,the transfection efficiency was detected by fluorescent microscope.The cell vitality was measured by typan blue stainning.Results The transfection efficiency was the highest and the cell vitality was not different from other subgroups when ultrasound was radiated at intensity of 1.0 W/cm~2 for 30 s.The transfection efficiency of plasmid-microbubble-ultrasound group were statistically higher than that of other groups(P<0.05).Conclusions Under optimal sound intensity and exposure time,ultrasound-mediated microbubble destruction could enhance the gene transfection in NSCs.更多还原