提高多药耐药基因导入人CD_(34)~ 细胞转导效率的探讨

目的　探讨逆转录病毒介导的多药耐药基因 (mdr1)导入人CD34 细胞的影响因素 ,以提高基因转导效率。方法　用流式细胞术 (FCM)检测不同组合细胞因子及人骨髓基质细胞 细胞因子支持的基因转导效率 ;用造血祖细胞集落培养观察耐药性 ;用FCM检测不同浓度紫杉醇素对基因转导细胞的作用。结果　细胞因子SCF Flt配体 (FL) IL 3组合支持的基因转导效率高于其它组合 (SCF IL 6 IL 3,SCF IL 6 IL 3 Tpo ,SCF IL 3)。骨髓基质细胞 细胞因子 (SCF FL IL 3)支持的基因转导效率 (2 0 .5 % )又高于单纯用该组细胞因子的转导效率 (15 .2 % ) ,并且前者形成的抗性集落形成细胞数多于后者。在 10ng ml紫杉醇作用下基因转导CD34 细胞的阳性率可达 38.5 %。结论　骨髓基质细胞 细胞因子 (SCF FL IL 3)对基因转导有较强的促进作用 ;一定浓度的紫杉醇具有富集基因转导细胞的作用

Exploration of factors increasing transfer efficiency of retroviral mediated multidrug resistance gene (mdr1) into human CD_(34)~ cells

Objective To explore factors influencing transfer of retroviral mediated multidrug resistance gene (mdr1) into human CD 34 cells. Methods Transduction efficiency in the presence of different combinations of cytokines and human bone marrow stromal cells plus cytokines were determined by flow cytometry (FCM).Drug resistance was evaluated by plating yields of cultured hematopoietic progenitor cells. The effect of Taxol at different concentrations on mdr1 gene transfer cells was determined by FCM. Result Transduction efficiency in the presence of SCF FL IL 3 was higher than that in other combinations of cytokines(SCF IL 6 IL 3,SCF IL 6 IL 3 Tpo,SCF IL 3). Transduction efficiency (20.5%) in the presence of bone marrow stromal cells plus cytokines(SCF FL IL 3) was higher than that (15.2%) without stromal cells, and the number of drug resistant colony forming cells was more in the former than in the latter. The percentage of CD 34 cell with the gene transduction at 10?ng/ml Taxol reached 38.5%. Conclusion Human bone marrow stromal cells plus cytokines (SCF FL IL 3) is more effective in enhancing mdr1 gene transduction. Taxol at a certain concentration can enrich mdr1 gene transferred cells.