活细胞标记和流式细胞术应用于体外检测巨噬细胞吞噬活性的方法建立  

A method to explore phagocytosis activities of macrophages in vitro by cell tracing and flow cytometry

作　　者：王佳佳[1] 王磊[1,2] 宋兴辉[1] 李艳伟[1] 郭春[1] 黄莹莹 刘丽[1] WANG Jia-Jia;WANG Lei;SONG Xing-Hui;LI Yan-Wei;GUO Chun;HUANG Ying-Ying;LIU Li(Core Facilities,Zhejiang University School of Medicine,Hangzhou 310058,China)

机构地区：[1]浙江大学医学院公共技术平台,杭州310058 [2]焦作师范高等专科学校,焦作454000

出　　处：《中国免疫学杂志》2020年第6期714-718,共5页Chinese Journal of Immunology

基　　金：浙江大学实验技术研究项目(SYB201610);浙江省教育厅一般科研项目(Y201942279)资助。

摘　　要：目的:通过活细胞标记和流式细胞术的联合应用,建立一种方法研究体外巨噬细胞的吞噬活性。方法:研究采用活细胞示踪染料CFSE标记对照组巨噬细胞,对未标记CFSE的细胞使用免疫刺激剂LPS处理,两组细胞混匀后与预先超声处理过的Alexa594标记的大肠杆菌孵育,在一个体系中完成吞噬过程,流式细胞术检测双荧光信号来评价对照组细胞和LPS处理后细胞的吞噬活性。结果:对照组和LPS处理组巨噬细胞的吞噬率分别为48.6%和62.3%,LPS处理后巨噬细胞的吞噬活性明显升高,与文献报道一致。实验表明,通过活细胞标记区分,在同一体系中完成LPS处理和未处理细胞的吞噬活性检测是非常可行的。也正因为待比较吞噬活性的两组细胞处在同一个吞噬环境中,吞噬率反映的吞噬活性更为准确。结论:本研究应用活细胞标记和流式细胞术,成功建立了一种实现体外处理的巨噬细胞吞噬活性的检测方法,简便、准确、可信度高。Objective:To establish a method for evaluating macrophage′s phagocytic activities in vitro by cell tracing and flow cytometry.Methods:One group of macrophages set as the control was labeled with cell tracing dye CFSE.The other group unlabeled with CFSE was treated with LPS.The two groups of macrophages were mixed and then incubated with the sonication-treated E.coli labeled with Alexa594 to enable the uptake of E.coli by the cells.The phagocytotic activities of macrophages were finally determined by flow cytometry for the presence of indicated signals.Results:The percentage of phagocytic cells as determined by flow cytometry in the control group and LPS treated group was 48.6%and 62.3%,respectively.The phagocytic activities of macrophage increased after treatment of LPS,which was consistent with previous research.Therefore,the results indicate the employment of our method for evaluating phagocytic activities of macrophage was feasible.Moreover,it was more accurate and convincing for the analysis of phagocytosis for two group cells in one homogeneous system.Conclusion:We successfully established a method of evaluating the phagocytic activities of macrophages in vitro by cell tracing and flow cytometry,which is feasible,accurate and convincing.

关 键 词：巨噬细胞 吞噬 细胞示踪染料 流式细胞术 

分 类 号：R392-3[医药卫生—免疫学]