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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
Objective To investigate the influencing factors of retrovirus-mediated multidrug resistance gene (mdr1) transfection into human CD34 + cells and to improve the efficiency of gene transduction. Methods Flow cytometry (FCM) was used to detect the gene transduction efficiency of different combinations of cytokines and human bone marrow stromal cells + cytokines. The hematopoietic progenitor cell colonies were used to observe the drug resistance. FCM was used to detect the effect of different concentrations of paclitaxel on gene transfer Guide the role of cells. Results The gene transduction efficiency of SCF + Flt ligand (FL) + IL 3 combination was higher than that of other combinations (SCF + IL 6 + IL 3, SCF + IL 6 + IL 3 + Tpo, SCF + IL 3). The transduction efficiency (20.5%) of BMSCs + cytokines (SCF + FL + IL 3) was higher than that of transduction of BMSCs alone (15.2%), and the former formed The number of resistant colony-forming cells was greater than the latter. The positive rate of CD34 + cells transduced with 10ng ml paclitaxel was 38.5%. Conclusion BMSCs + cytokines (SCF + FL + IL 3) have a strong role in gene transduction; paclitaxel with a certain concentration has the effect of enriching gene transduction cells