流式细胞术纯化脐血嗜碱性粒细胞  

Flow cytometry in purification of umbilical cord blood-derived basophils

作　　者：杨玲[1] 许以平[2] 姚苏杭[2] 熊瑛[3] 祝捷[4] 王克敏[5] 郭胤仕[2] 

机构地区：[1]上海交通大学医学院附属新华医院呼吸科,200092 [2]上海交通大学医学院附属仁济医院过敏中心 [3]上海交通大学医学院附属新华医院妇产科,200092 [4]上海交通大学医学院附属仁济医院妇产科 [5]上海交通大学医学院生化教研室

出　　处：《上海医学》2010年第1期75-77,共3页Shanghai Medical Journal

摘　　要：目的采用流式细胞术提纯脐血嗜碱性粒细胞,以期得到研究过敏性疾病发病机制的理想标本。方法6份足月产妇新生儿脐血经沉淀红细胞及Ficoll-泛影葡胺处理后,采用流式细胞术分选CD203c-PE+、CD45-PerCPint的细胞。通过特异性识别嗜碱性粒细胞的阿利新蓝染色和细胞计数脐血嗜碱性粒细胞的纯度和回收率。结果流式细胞术分选嗜碱性粒细胞所得的纯度为(95.02±2.94)%,回收率为(61.42±5.95)%。结论用流式细胞术分选脐血中CD203c-PE+、CD45-PerCPint的细胞可获得高纯度、高回收率的嗜碱性粒细胞。Objective To use flow cytometry for purification of human basophils from umbilical cord blood.Methods Umbilical cord blood samples were collected from 6 healthy,full-term deliveries.After separating the red blood cells by precipitation,the mononuclear cells were isolated by Ficoll-Hypaque.Then flow cytometry（FCM） was used to sort CD203c-PE^＋ and CD45-PerCP^int cells.The purity and recovery of basophils were examined by Alcian blue staining and cell counting.Results The purity and recovery of basophils by FCM were（95.02±2.94）% and（61.42±5.95）%,respectively.Conclusion Cell sorting for CD203c-PE^＋,CD45-PerCP^int cells by FCM is a better method to obtain pure umbilical cord blood-derived basophils,with high purity and recovery.

关 键 词：流式细胞术 免疫磁珠 细胞纯化 脐带血 嗜碱性粒细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]