一种低成本的逆转录病毒感染白血病细胞的方法探析

A low cost method of retrovirus infection in leukemic cells

为了降低实验成本,解决悬浮细胞难转染问题,本研究进行逆转录病毒包装及感染悬浮细胞—白血病细胞的实验。采用磷酸钙转染方法将逆转录病毒质粒MSCV-PIG、Gag-pol与VSVG共转染入293T细胞,通过旋转法将病毒原液与KG-1a白血病细胞共孵育,最后用嘌呤霉素筛选稳定细胞株。逆转录病毒原液感染KG-1a细胞48 h后KG-1a细胞的感染效率约为10%。但嘌呤霉素稳筛后,携带逆转录病毒载体的KG-1a细胞阳性率可达85%以上。通过低成本的磷酸钙转染方法获取病毒,采用病毒原液成功感染较难转染的白血病细胞,为做基础研究的科研人员提供借鉴。

To reduce experimental cost and solve the problem for hard to transfect vectors in suspension cells,this study takes an experiment for packaging retrovirus and infecting suspension cells such as leukemic cells. The retroviral vectors MSCV-PIG,Gag-pol and VSVG were transfected into 293 T cells by calcium phosphate transfection method. Then,the unconcentrated viral liquids were incubated with KG-1 a leukemic cells by centrifugation. At last,puromycin was used to screen stable cell lines. Infection of KG-1 a cells with retrovirus stock solution for 48 h,the efficiency of infection was about 10%. However,the percentage of KG-1 a cells which successfully infected by retrovirus was more than 85% after puromycin screening. The retrovirus is packaged by the low-cost calcium phosphate transfection method and the hard-to-transfected leukemic cells are successfully infected by unconcentrated viral liquids,which offers reference to fundamental researchers.