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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

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【作者】 伊焕发唐岩甄昱唐军民孙红亚

【Author】 YI Huan fa 1, TANG Yan 2, ZHEN Yu 2, TANG Jun min 2, SUN Hong ya 2 (1.Department of Anatomy, Dalian Medical University, Dalian 116027; 2.Department of Histology and Embryology, Beijing University Health Science Center, Beijing 100083, China)

【机构】 大连医科大学解剖学教研室北京大学医学部组织与胚胎学教研室北京大学医学部组织与胚胎学教研室 辽宁大连116027北京100083北京100083

【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 ,为进一步进行DC与肿瘤的融合实验及临床应用提供了可能

【Abstract】 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.

【基金】 国家自然科学基金资助项目 (NO 3 9970 3 80 )
  • 【文献出处】 解剖科学进展 ,Progress of Anatomical Sciences , 编辑部邮箱 ,2003年03期
  • 【分类号】R392
  • 【下载频次】107
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