三种分离人外周血单核细胞方法的比较  

Comparison of three methods of separating human peripheral blood mononuclear cells

作　　者：陈丹[1] 王小东[2] 童静植 丁文[2] 孙科[2] 何景华[1] 强兆艳[1] 李光[3] 康毅[1] 

机构地区：[1]天津医科大学基础医学院药理学教研室,天津300070 [2]天津医科大学 [3]天津医科大学基础医学院生物学教研室,天津300070

出　　处：《天津医科大学学报》2014年第6期483-485,共3页Journal of Tianjin Medical University

基　　金：国家高技术研究发展计划(863计划)第八课题组子课题(2012AA021003)

摘　　要：目的:用3种方法从人外周血分离单核细胞,比较细胞纯度、得率、实验成本及所需时间,选择最适宜的分离方法。方法:用密度梯度离心法获得外周血的单个核细胞,分别用贴壁法、免疫磁珠法和流式细胞术法进一步分离单核细胞。经CD14抗体标记,用流式细胞术检测细胞纯度,计算细胞得率、实验成本及所需时间。结果:用贴壁法、免疫磁珠法和流式细胞术法分离单核细胞,细胞纯度分别为18.8%、80%和73.4%;细胞得率为5%、11%和7.5%;每获得2×107个单核细胞,实验成本约500元、1 500元和2 100元,实验耗时约6 h、6 h和10 h。结论:免疫磁珠法是最适宜的分离人外周血单核细胞的方法,具有细胞纯度与得率较高,实验成本较低,耗时较短的优点。Objective： To separate monocyte from human peripheral blood based on cell purity, cell yield rate, experiment cost and the time required. Methods： Peripheral blood mononuclear cells were obtained via density gradient centrifugation method. Then the adherent method, immune magnetic beads method and flow cytometry method were used to separate monocyte from peripheral blood mononuclear cells. Cell purity was detected by flow cytometry after being marked with CD14 monoclonal antibody. Cell yield rate, experiment cost and the time required were calculated. Results： The cells purity was 18.8% by adherent method while cells purity was 80% and 73.4% respectively indicated by immune magnetic beads and flow cytometry methods. The cell yield rates were 5%, 11% and 7.5%, respectively. The experiment costs were about 500 yuan, 1 500 yuan and 2100 yuan every 2×10^7 mononuclear cells while the time required for experiment were about 6 h, 6 h and 10 h, respectively. Conclusion： Immune magnetic beads method is the most appropriate method to separate monocyte, with the advantages of higher cell purity and yield rate, lower experiment cost and shorter time as compared with other methods.

关 键 词：单核细胞 贴壁法 免疫磁珠 流式细胞术 

分 类 号：R446[医药卫生—诊断学]