外周血嗜碱粒细胞磁珠分选法分选及意义  

Clinical significance of the magnetic activated cell sorting of peripheral blood basophils

作　　者：孙仁山[1] 隋建峰[2] 冉新泽[3] 唐书谦[1] 杨涛[1] 杨宏珍[1] 李凤[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所皮肤科,重庆400042 [2]第三军医大学基础部医学试验中心,400038 [3]第三军医大学预防医学院复合伤研究所,重庆400038

出　　处：《重庆医学》2010年第23期3224-3225,共2页Chongqing medicine

基　　金：国家自然科学基金资助项目(30972647);第三军医大学科研基金资助项目(2008XG179)

摘　　要：目的探讨磁珠分选法(MACS)负性筛选方法获得高纯度嗜碱粒细胞用于速发型过敏反应的研究。方法以Ficoll密度梯度离心分离细胞。用活性抗体标记的免疫磁珠进行MACS分选,用激光共聚焦方法鉴定嗜碱粒细胞活性。结果 Ficoll密度离心后,获得嗜碱粒细胞的纯度为(12.6±3.87)%,MACS纯化后,其纯度可提高到(95.6±3.96)%,细胞活性为(98.6%±0.83)%。激光共聚焦证实,纯化后的嗜碱粒细胞活化后CD63表达增强,细胞功能完整。结论用MACS负性筛选方法可获得高纯度的嗜碱粒细胞,细胞功能完整,可用于速发型过敏反应的研究。Objective To obtain pure basophils form whole blood by negative MACS（Magnetic activated cell sorting）for hypersensitive reaction study.Methods The mononulear cells were discarded after Ficoll gradient centrifuge.The basophils were sorted from the rest white blood cells by the method of MACS and the purity rate after sorting was analyzed.The CD63 expression on basophils were observed with laser confocal microscopy.Results The purity of the basophils was 95.6%±3.96% and live cell rate was 98.6%±0.83% after the MACS.The CD63 expression on basophil surface was greatly increased after cell activations.Conclusion The protocols in our study for isolating and enriching basophils from the whole blood by Ficoll isolation and MACS negative selection proccess can acquire basophils with high abundance,enrichment and viability and the cells can be utilized in hypersensitive reaction studies.

关 键 词：嗜碱粒细胞 负性筛选 激光共聚焦显微镜 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]