人工合成的siRNA转染外周血活化T淋巴细胞方法学研究  

The methodology for transfection of human peripheral blood activated T lymphocytes with synthetic siRNA.

作　　者：葛晓松[1] 薛祝平[1] 李柏青[1] 

机构地区：[1]蚌埠医学院免疫学教研室安徽省感染与免疫重点实验室,蚌埠233030

出　　处：《生物学杂志》2009年第1期79-80,95,共3页Journal of Biology

基　　金：安徽省自然基金资助项目(050430603)

摘　　要：观察人工合成siRNA与脂质体DMRIE-C Reagent的复合物对人外周血活化T淋巴细胞转染效果。人外周血标本中分离的外周血单个核细胞用抗原和IL-2刺激扩增培养,产生活化的T淋巴细胞,用不同配比浓度的荧光标记的siRNA-FAM与脂质体的复合物转染活化后的T淋巴细胞,分别采用转染一次和连续转染两次的方法,并在转染后4h,分别取样本,通过流式细胞测定技术,比较两种方法的转染效率,并筛选出最佳转染效率剂量配比。活化T细胞用转染一次和两次的方法后,平均转染阳性率分别为(12.5±0.64)%和(50.37±6.77)%。采用连续两次转染的方法进行转染,转染效率要明显高于一次转染法。The effect of siRNA/DMERI-C Reagent complex on transfection of human activated T lymphocytes from peripherial blood was investigated. The peripheral blood mononuclear cells （PBMC） was isolated from blood by Ficoll-hypaque method, and treated with IL-2 and antigen to produce activated T cells. Different finial concentration of siRNA-FAM was transfectated into activated T cells by DMERI-C Reagent for one or two times . 4h later, the sample was analgzed by FACS Calibur. The result showed the percentages of siRNA-FAM in activated T cells were （ 12. 5 ± 0. 64） % and （50. 37 ± 6. 77 % at average （ n = 4） for transfecting once and twice respectively. Use the method transfection by two times can induce the siRNA into activated T cells more successful than that by one times.

关 键 词：SIRNA 转染 脂质体 

分 类 号：Q813[生物学—生物工程]