不同细胞因子对培养人外周血树突状细胞的影响  

EFFECTS OF DIFFERENT CYTOKINES TO HUMAN BLOOD DENDRITIC CELLS

作　　者：张宏[1] 钟翠平[1] 成令忠[1] 顾云娣[1] 叶胜龙[1] 

机构地区：[1]上海医科大学基础医学院组胚学教研室

出　　处：《上海医科大学学报》1998年第3期215-218,共4页Journal of Fudan University(Medical Science)

摘　　要：目的建立从人外周血分离、纯化、培养、扩增树突状细胞（ＤＣ）前体的方法，研究细胞因子对ＤＣ体外增殖、分化成熟的影响。方法人外周血经血细胞分离仪及Ｆｉｃｏｌ、Ｐｅｒｃｏｌ等不连续密度梯度离心获得的含ＤＣ前体细胞组分用重组人粒细胞／巨噬细胞集落刺激因子（ｈｒＧＭ－ＣＳＦ）培养或用ＧＭ－ＣＳＦ及白细胞介素－４（ＩＬ－４）联合培养；光镜及电镜观察及ＡＢＣ法免疫细胞化学染色。结果分离的ＤＣ前体经１周左右时间培养，细胞数量可扩增２０～３０倍，纯度达９０％以上，ＧＭ－ＣＳＦ与ＩＬ－４联合培养所得到的ＤＣ数量约为用ＧＭ－ＣＳＦ培养的１．５倍。ＤＣ具有典型的树枝状或裙褶状突起，并表达高水平的ＨＬＡ－ＤＲ，其ＣＤ１４、ＣＤ１９、ＣＤ３的表达均为阴性。结论用ＧＭ－ＣＳＦ和ＩＬ－４联合作用更能促进ＤＣ的体外扩增及分化。URPOSE To establish the methods of isolation, purification and propagation of dendritic cell (DC) precursor from human peripheral blood and to study the effects of different cytokines on human blood DCs.METHODS DC precursors were isolated by discontinuous density gradient centrifugation, and cultured with human recombinant granulocyte/macrophage colonystimulating factor (hr GM-CSF) on plus human recombinant interleukin 4( hr IL-4). The morphologic features of DCs were observed by light and electron microscopes. Cell phenotypes were detected by ABC immunocytochemical staining.RESULTS The number of DCs was 20 to 30 folds increased and the purity of cultured DC was over 90%. Cell number in the GM-CSF plus IL-4 group was 1.5 times as many as that in the GM-SCF group. Cultured cells in these two groups exhibited special dendritic morphology of the mature DC and expressed HLA-DR highly, while CD14, CD19 and CD3 expressions were negative.CONCLUSIONS The results indicate that a combination of GM-CSF plus IL-4 could greatly promote the proliferation and differentiation of DCs in vitro.

关 键 词：树突状细胞 细胞培养 细胞因子 细胞表型 

分 类 号：R329－33[医药卫生—人体解剖和组织胚胎学]