MACS法和EasySep法分选人外周血调节性T细胞的效果比较  

Comparison of MACS and EasySep Methods for Sorting Human Peripheral Blood Regulatory T Cells

作　　者：武星星 孙强 阳玉洁 王钦 魏玉惠 王云甫 WU Xing-xing;SUN Qiang;YANG Yu-jie;WANG Qin;WEI Yu-hui;WANG Yun-fu(Department of Neurology,Taihe Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China;Department of Neurology,Xiaogan Central Hospi-tal,Xiaogan,Hubei 432000,China;Postgraduate Training Basement of Jinzhou Medicical University,Taihe Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]十堰市太和医院·湖北医药学院附属医院神经内科,湖北十堰442000 [2]孝感市中心医院神经内科,湖北孝感432000 [3]锦州医科大学十堰市太和医院研究生培养基地·湖北医药学院附属医院,湖北十堰442000

出　　处：《湖北医药学院学报》2020年第4期334-338,F0003,共6页Journal of Hubei University of Medicine

基　　金：国家自然科学基金资助项目(81672138);湖北医药学院研究生科技创新项目(YC2019033)。

摘　　要：目的:比较MACS法与EasySep法两种分选人外周血调节性T细胞(regulatory T cells,Tregs)效果的差异,探索一种更高效分选Tregs的方法。方法:采集健康成年人的外周血,用Ficoll密度梯度离心法分离出人外周血单个核细胞(Peripheral blood mononuclear cells,PBMC),将PBMC均分为两组:MACS法分选组和EasySep法分选组,分别选用基于分离柱的人CD4^+CD25^+调节性T细胞MACSTM分选试剂盒和EasySepTM人CD4^+CD25high T细胞分选试剂盒两种分选方法分离得到Tregs,并对其数量、存活率、细胞形态和纯度等进行比较。结果:MACS法分选组的Tregs细胞数为:(2.19±0.21)×10^6个,EasySep法分选组的细胞数为:(1.73±0.12)×10^6个,明显低于MACS法分选组(t=4.551,P<0.05);MACS法分选组细胞存活率为:(94.37±4.15)%,EasySep法分选组细胞存活率为(91.98±1.23)%,明显低于MACS法分选组(t=3.073,P<0.05);MACS法分选组细胞培养后部分细胞形态不规则,为杆状或椭圆状,而EasySep法分选组细胞形态保持完好,无变形;MACS法分选组的CD4^+CD25^+Treg比例为:(96.52±1.42)%,EasySep法分选组为:(85.67±10.58)%,明显低于MACS法分选组(t=2.489,P<0.05);MACS法分选组CD4^+CD25^+CD127low/-Tregs的比例为:(67.85±1.11)%,EasySep法分选组为:(63.08±1.11)%,明显低于MACS法分选组(t=7.448,P<0.05)。结论:两种方法各有优势,但基于分离柱的MACS法得到的Tregs数量多,纯度高,且具有功能性的Tregs比例高,是一种更为高效的分选人外周血调节性T细胞的方法。Objective To compare the difference between MACS and EasySep technologies in sorting human peripheral blood regulatory T cells(Tregs),and to explore a more efficient method for Tregs sorting.Methods Peripheral blood mononuclear cells(PBMCs)were isolated from healthy adults using Ficoll density gradient centrifugation.The PBMCs were divided into two groups:group MACS and group EasySep.Tregs were isolated by two sorting methods:human CD4^+CD25^+regulatory T cell MACSTM sorting kit based on separation column and EasySepTM human CD4^+CD25highT cell sorting kit,and their num⁃ber,survival rate,cell morphology and purity were compared.Results The number of Tregs cells sorted in group MACS was(2.19±0.21)×10^6,and that in group EasySep was(1.73±0.12)×10^6,(t=4.551,P<0.05).The survival rate of group MACS was(94.37±4.15)%,and the survival rate of EasySep group was(91.98±1.23)%,which was significantly lower than that of group MACS(t=3.073,P<0.05).Some cells in the group MACS had irregular shapes after cell culture and were rod-shaped or elliptical,while the cell morphology of the group EasySep remained intact and no deformation.The pro⁃portion of CD4^+CD25^+Tregs in group MACS was(96.52±1.42)%,in group EasySep was(85.67±10.58)%,The propor⁃tion of CD4^+CD25^+CD127low/-Tregs in group MACS was(67.85±1.11)%,and that in group EasySep was(63.08±1.11)%,which was significantly lower than that of group MACS(t=7.448,P<0.05).Conclusion The two methods have their own advantages,but the MACS method based on separation column has a large number of Tregs,high purity and high proportion of functional Tregs,so it is a more efficient method for sorting human peripheral blood regulatory T cells.

关 键 词：调节性T细胞 免疫磁珠分选 

分 类 号：R392[医药卫生—免疫学]