Lentivirus载体对许旺细胞的转染效率

背景：近年来研究较多的LV载体,因其强大的转染能力以及较高的转染效率等特点,在基因转染的实验中应用越来越广泛。目的：进一步验证Lentivirus载体在体外对原代许旺细胞的转染效率。方法：以Lentivirus三质粒系统构建病毒载体,在体外分别以MOI值为1,5,10,20,40对原代大鼠许旺细胞进行转染,转染后第1,3,5,7,9天在荧光显微镜下观察Lentivirus携带的荧光表达情况,并在显微镜的计数方格内计算细胞的转染情况。发出绿色荧光的为转染成功的许旺细胞,否则认为没有转染病毒载体的,从而算出转染效率。结果与结论：在病毒转染3 d后,在各不同MOI值的培养孔中,均能观察到极少量的荧光反应,在第5天时,荧光数量较前明显增多。第7天时达到高峰,第9天的荧光数量与第7天变化不明显。从细胞转染效率来看,不同的MOI值明显不同,MOI值为1时的转染效率约为45%,MOI值为5时为80%,MOI值为10时为90%,MOI值为20时为78%,MOI值为40时转染效率为70%。

Transfection efficiency of Lentivirus carrier to Schwann cellsin vitro

BACKGROUND： The Lentivirus carrier studied in recent years has been widely used in the gene transfection experiment due to its strong transfection ability and high transfection efficiency. OBJECTIVE To study the transfection efficiency of Lentivirus carrier to primary Schwann calls in vitro. METHODS： The virus vector was constructed with Lentivirus three-plasmid system. In vitro, the pdmary Schwann cells were transfected by recombinant virus with different-multiplicities of infection （1, 5, 10, 20 and 40）. At 1, 3, 5, 7 and 9 days after transfection, fluorescent expression of Lentivirus was observed under fluorescence microscope, and the transfection efficiency was calculated in the counting squares of the microscope. Green fluorescence presented the transfected Schwann cells, and the others were untransfected Schwann cells. Then the transfection efficiency was calculated. RESULTS AND CONCLUSION： At 3 days after transfection, the small amount of fluorescence could be seen in the culture dishes with different multiplicities of infection, and then the fluorescence amount was increased at 5 days after transfection. It reached peak after 7 days. There was no significant difference in the fluorescence amount between 7 and 9 days after transfection. Different multiplicities of infection could lead to different transfection efficiencies： the transfection efficiency was 45% when the multiplicity of infection was 1; the transfection efficiency was 80% when the multiplicity of infection was 5; the transfection efficiency was 90% when the multiplicity of infection was 10; the transfection efficiency was 78% when the multiplicity of infection was 20; and the transfection efficiency was 70% when the multiplicity of infection was 40.