细胞因子对逆转录病毒载体介导的小鼠骨髓造血干细胞基因转移效率的影响

本实验探讨细胞因子SCF,IL-3和IL-6对基因转移效率的影响,为临床基因治疗提供可靠依据。以小鼠造血干细胞为靶细胞,应用包装细胞株PA317-GCGPXSN制备病毒上清,实施基因转移,采用FACS,GM-CFU,PCR和Southern blot方法测定基因转移效率。实验结果为:SCF,IL-3,IL-6单用或联合应用后,FACS测定的基因转移效率为0.07％-0.20％,GM-CFU测定的结果为20.4％-46.40％,阳性对照组测定的结果则分别为0.06％和10.92％。结论提示SCF/IL-3,SCF/IL-3/IL-6联合应用能显著提高基因转移效率。

Study on effect of cytokines on gene transfer efficiency in mice bone marrow hematopoietic stem cells mediated by retroviral vector

The effect of cytokines SCF, IL-3 and IL-6 on gene transfer efficiency in mice bone marrow hematopoietic stem cells, mediated by retroviral vector, has been studed. Viral supernatants were harvested with PA317-GCGPXSN, and then made gene transfer, using mice hematopoietic stem cells as target cell. The efficiency of gene transfer was determined by FACS, CFU-GM, PCR and Southern blot. Results showed that the efficinecy of gene transfer using SCF, IL-3, IL-6 alone or in combination were 0.07% -0.20% (FACS) and 20.45% -46.40% (CFU-GM), but the efficiency of positive control groups was 0.06% and 10.92%, respectively. It is concluded that SCF/IL-3/IL-6 and SCF/IL-3 could obviously increase the efficiency of gene transfer into hematopoietic stem cells mediated by retroviral vector.