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利用磁性细胞分选技术阴性选择法分离外周血嗜酸性粒细胞 |
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李一荣 吴健民 邹萍
摘 要 目的：从健康献血者和哮喘等变态反应性疾病病人的外周血中分离出高纯度、高存活率的嗜酸性粒细胞。方法：采用磁性细胞分选技术的阴性选择法分离外周血嗜酸性粒细胞，并检测分离后嗜酸性粒细胞的纯度、存活率，计算其回收率。结果：①健康献血者外周血嗜酸性粒细胞的回收率为69.7%±6.8%，存活率95.4%±1.8%，纯度为90.7%±4.3%；②哮喘等变态反应性疾病病人外周血嗜酸性粒细胞的回收率为60.4%±5.4%，存活率为96.1%±0.7%，纯度为92.6%±5.1%。结论：采用磁性细胞分选技术的阴性选择法，能从外周血中分离出高纯度、高存活率的嗜酸性粒细胞，因此该方法是一种高效的分离外周血嗜酸性粒细胞的免疫学方法。
关键词 磁性微球 哮喘 嗜酸性粒细胞
中国图书分类号 R392.33
Isolation of eosinophils from peripheral blood by the negative selection procedure of magnetic activated cell sorting
LI Yi-Rong，WU Jian-Min，ZOU Ping.
Laboratory Department，Union Hospital，Tong Ji Medical University，Wuhan，430022
Abstract Objective:To isolate eosinophils of high purity and viability from peripheral blood of healthy donors and patients with allergic disease(such as asthma).Methods:Eosinophils of peripheral blood were isolated by the negative selection procedure of magnetic activated cell sorting and their purity and the ratio of viability and recovery were assessed.Results:(1)The purity and the ratio of viability and recovery of eosinophils from healthy donors were 90.7%±4.3%，95.4%±1.8% and 69.7%±6.8% respectively.(2)The purity and the ratio of viability and recovery of eosinophils form patients with allergic disease (such as asthma) were 92.6%±5.1%，96.1%±0.7% and 60.4%±5.4% respectively.Conclusion:By the negative selection procedure of magnetic activated cell sorting，eosinophils of high purity and viability were acquired from peripheral blood.So the procedure is a highly effective immunological method which used to isolate eosinophils.
Key words Magnetic microbead Asthma Eosinophil
长期以来，嗜酸性粒细胞的分离多采用不连续的密度梯度离心法，但是由于外周血中嗜酸性粒细胞的含量较少，约为白细胞的0.5%～5%，而且其密度范围与中性粒细胞部分重叠，因此高度纯化外周血中嗜酸性粒细胞有一定的难度。1979年，Vadas采用高渗的Metrizamide进行密度梯度离心分离外周血中嗜酸性粒细胞［1］，随后Gartner将高渗的Metrizamide换成等渗Percoll液分离外周血嗜酸性粒细胞［2］，尽管能够达到一定的纯度和回收率，但是Metrizam[1] [2] 下一页
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