人外周血树突状细胞的诱导与鉴定  

Induction and identification of dendritic cells from human peripheral blood monocytes

作　　者：侯治富[1] 郭楠[2] 高申 郑德明[1] 卜丽莎[1] 高嵩[3] 才华[1] 

机构地区：[1]吉林大学中日联谊医院中心研究室,吉林长春130033 [2]北京中医药大学附属东直门医院检验科,北京100001 [3]浙江大学附属第二医院检验科,浙江杭州310009

出　　处：《吉林大学学报（医学版）》2005年第5期657-660,F0002,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30471610);吉林大学创新基金项目(403010123138)

摘　　要：目的:体外诱导、培养人外周血单核细胞获得不同成熟阶段的树突状细胞(DCs)。方法:用贴壁法从健康人外周血浓缩白细胞获取单核细胞,第一阶段在GM-CSF+IL-4存在的条件下培养7 d,获得未成熟DCs;第二阶段在GM-CSF+TNF-α联合诱导下培养至14 d,获得成熟的DCs。对DCs的形态进行显微镜下观察,用流式细胞仪检测其表型,用MTT的方法对DCs的功能进行检测。结果:获得的未成熟DCs中度表达CD1a、共刺激分子,高表达HLA-DR分子,在DCs的成熟期共刺激分子、HLA-DR及CD83、CD25分子均高度表达,刺激同种异型T淋巴细胞增殖的能力强。结论:成功地建立了诱导人外周血单核细胞获得不同发育阶段DCs的方法,并获得了大量纯度较高的DCs。Objective To induce and culture purity dendritic cells （DCs） of different maturation phase from human peripheral blood monocytes in vitro. Methods The monocytes were acquired from human peripheral blood mononuclear cells （PBMC） by adhesion method. The first step was to work on a 7-day culture of monocytes in medium supplement with GM-CSF and IL-4, and the cells were actually immature. In the second step, the cells were cultured in the medium supplement with GM-CSF and TNF-α, and cultured until the 14th day. Mature DCs were obtained. Cells harvested were identified for their morphology by microscope, properties of DCs detected by FCM, function with MTT method. Results Immature dendritic cells expressed CDla molecules and costimulating molecules in middle level, HLA-DR in high level. In maturation phase DCs expressed costimulating molecules, HLA-DR molecules, CD83, and CD25 highly. These DCs could stimulate proliferation of allogenic T lymphocytes and increase killing ability of CTL activated by DCs. Conclusion The method to obtain DCs of different maturation phases from human peripheral blood monocytes is successfully set up. A number of DCs with high purity are obtained.

关 键 词：树突细胞 细胞 培养的 诱导 共刺激分子 

分 类 号：R392.12[医药卫生—免疫学]