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  

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

作　　者：王成国[1] 薛克营[1] 程立[1] 李威[1] 石明[1] 王正艳[1] 

机构地区：[1]郧阳医学院附属东风医院呼吸科,湖北十堰442008

出　　处：《郧阳医学院学报》2008年第4期295-297,共3页Journal of Yunyang Medical College

摘　　要：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。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 OVAimmune 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 carried out for detection. Results Before MACS enrichment, CD4^＋ CD25 ^＋ Treg,/CD4^＋ T ratio was 11.07 %, and CD4 ^＋ T/total cells ratio was 21.88%. After MACS negative enrichment, CD4^＋ CD25^＋ Treg,/CD4^＋ T ratio was 10.95%, and CD4 ^＋ T/total cells ratio was 89.28%. After MACS positive enrichment, CD4 ^＋ T/total cells ratio was over 95% ,CD4^＋ CD25 ^＋ Treg,/CD4 ^＋ T ratio was 90.92% ,and the purity of CD4 ^＋ CD25 ^＋ Treg was more than 90%. Conclusion High purity CD4^＋ CD25 ^＋ Treg were successfully isolated from PBMCs of OVA-immune tolerance rats by MACS.

关 键 词：CD4^+CD25^+ 调节性T细胞 磁激活细胞分选 外周血单个核细胞 

分 类 号：R392[医药卫生—免疫学]