人外周血CD4^+树突状细胞的纯化及鉴定  

Purification and Identification of CD4^+ Dendritic Cells from Human Peripheral Blood

作　　者：伊焕发[1] 唐岩[2] 甄昱[2] 唐军民[2] 孙红亚[2] 

机构地区：[1]大连医科大学解剖学教研室,辽宁大连116027 [2]北京大学医学部组织与胚胎学教研室,北京100083

出　　处：《解剖科学进展》2003年第3期193-196,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目 (NO 3 9970 3 80 )

摘　　要：目的　建立人外周血树突状细胞 (dendriticcell,DC)的分离方法 ,观察其形态学和免疫组织化学特点 ,为下一步细胞融合提供DC来源。方法　以免疫磁珠分选法从人外周血单个核细胞中分离CD4 + DC ,流式细胞仪检测所得细胞的纯度 ,光镜、电镜和激光共聚焦扫描显微镜观察其形态 ,SP免疫细胞化学方法检测DC的分子表达。结果　此纯化方法所得细胞纯度可达到 80 %以上 ,形态学观察可见纯化细胞具有典型的DC特征 ,该细胞能高表达HLA DR和S 10 0分子。结论　免疫磁法可获得较高纯度典型DC 。Objective To purify and culture CD4 + dendritic cell (DC) from human peripheral blood in vitro to investigate the morphological properties and antigen expression of DC. Methods CD4 +DCs were isolated from human peripheral blood mononuclear cells (PBMC) by magnetic cell sorting (MACS). The purity of DC was analysed by fluorescence activated cell sorting (FACS) and the morphology was observed by photomicroscope, electron microscope and confocal analysis. The antigens were analysed by SP immunocytochemical method.Results The purity of DC was up to over 80% by MACS, the purified cells possessed typical DC morphological characters and DC expressed high levels of HLA DR and S 100 protein.Conclusion The method of MACS was highly effective for DC purification, and typical DCs were obtained from human peripheral blood.

关 键 词：外周血 树突状细胞 纯化 鉴定 细胞形态学 免疫组织化学 肿瘤细胞 

分 类 号：R730.3[医药卫生—肿瘤]