磁激活分选法双重分离大鼠外周血单个核细胞CD4~+CD25~+调节性T细胞

目的:应用磁激活细胞分选(MASC)技术从卵白蛋白(OVA)免疫耐受大鼠外周血单个核细胞(PBMCs)中分选出大量高纯度CD4+CD25+调节性T细胞(CD4+CD25+Treg),以研究其在支气管哮喘中的作用。方法:从OVA免疫耐受大鼠外周血中分离出PBMCs,应用MASC技术阴性分选CD4+T细胞,再阳性分选出CD4+CD25+Treg;流式细胞仪检测。结果:分选前的CD4+CD25+Treg细胞占CD4+T细胞数的11.07%,CD4+T细胞占总数的21.88%;CD4阴性分选后,CD4+CD25+Treg细胞占CD4+T细胞数的10.95%,CD4+T细胞占总数的89.28%;CD25阳性分选后,CD4+T细胞超过总数的95%,而CD4+CD25+Treg细胞占总数的90.92%,CD4+CD25+Treg细胞纯度大于90%。结论:应用MASC技术成功从OVA免疫耐受大鼠的PBMCs中分选出了高纯度的CD4+CD25+Treg细胞。

Isolation of CD4~+CD25~+ Regulatory T Cells by Negative and Positive Enrichment of Magnetic Activated Cell Sorter from rats PBMCs

Objective In order to study the possible roles of CD4+CD25+ regulatory T cells in the development of asthma,magnetic activated cell sorter(MACS)was used to isolate high purity CD4+CD25+ regulatory T cells(CD4+CD25+ Treg)from peripheral blood mononuclear cells(PBMCs)of OVA-immune tolerance rats.Methods PBMCs from blood of OVA-immune tolerance rats were isolated,CD4+ T cells were then purified by MACS negative enrichment,CD4+CD25+ Treg were further purified by MACS positive enrichment,flow cytometry was also ...