人外周血树突状细胞的体外培养及鉴定.pdfVIP

· 论 著· 一 —— _l_ 一 ～ 一 一 2007年12月第45卷第24期 人外周血树突状细胞的体外培养及鉴定冰 马国强 王佳烈 (内蒙古自治区医院呼吸病研究中心，呼和浩特010017) 【摘要】目的 在体外从人外周血中培养、鉴定树突状细胞。方法 取正常健康人新鲜外周血，经密度梯度离心法分离，获得单 个核细胞，取贴壁细胞加入重组人粒细胞一巨噬细胞集落刺激因子和重组人白细胞介素一4，体外培养第7天，加入肿瘤坏 死因子一 促进其分化成熟。用倒置显微镜观察其形态，用流式细胞仪进行表型测定。结果 从正常人外周血分离获得的单 个核细胞经rhGM—CSF，rhIL-4和TNF—CL联合培养，获得大量树突状细胞。具有典型的树突状细胞形态和免疫表型，高表 达HLA—DR，CD83，CD80，CD86、HLA—DR+CD83 及CD80 CD86 。结论 用rhGM—CSF，rhIL-4和TNF—CL联合培养可以从 人外周血诱导培养出大量的树突状细胞且高表达HLA—DR，CD80，CD83，CD86、HJ —DR+CD83+及CD80+CD86+。 键词】外周血；人类；树突状细胞；体外培养；鉴定 【中图分类号】R392．2 【文献标识码】A 【文章编号】1673—9701(2007)24_04—03 Culture and Identification of Dendritic Cells from Human Peripheral Blood in Vitro MA Guoqiang WANG Jialie Study Center of Respiratory Disease，Inner Mongolia Hospital，Huhehaote 010017 【Abstractl Objective To cultivate and identify dendritic eeHs from human peripheral blood in vitro．Methods e mononucIear ceHs were obtained from healthy human aseptic fresh peripheral blood by ficoH densi~ gradient eentrifugation．The adherent eeHs were cultured with human recombinant granuloeyte—macrophage colony-stimulating factor(rhGM-CSF)and human recombinant interleukin-4(rhI _4)． Tumor necrosis factor—CL(TNF—CL)was added in 7 days．Then the morphological features were observed by invert optical microscope，we evaluated the phenotypes changes by flow cytometry(FCM)．Results Large numbers of dendritic cells could be obtained from human p=iph— eral blood by combination of GM—CSF．II厂4 and TNF-CL．Dendritic ceHs had the typical morphology and phenotypes changes and highly暇一 pressed HLA—DR，CD80，CD83，CD86，HL 一DR CD83 and CD80+CD86~．Conclusion e results indicate that by comb ination of GM—CSF．II厂_4 and TNF— ，large numb ers ofdendritic cells could be obtained from human peripheral blood．Dendritic cells may typical lnor- phology and phenotypes chang