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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。
Objective To establish a method to separate, purify, culture and amplify dendritic cells (DC) precursors from human peripheral blood and to study the effect of cytokines on the proliferation, differentiation and maturation of DC in vitro. Methods The human DC-containing precursor cells were obtained by culturing human DCs with hrGM-CSF or transfected with GM-CSF And interleukin-4 (IL-4) were cultured with light microscope and electron microscope and ABC immunocytochemistry. Results The isolated DC precursors were cultured for about 1 week. The number of cells could be expanded by 20 ~ 30 times and the purity was over 90%. The amount of DC produced by co-culture of GM-CSF and IL-4 was approximately that of GM-CSF 1.5 times. DCs have typical dendritic or skirt pleat-like protrusions and express high levels of HLA-DR with negative expression of CD14, CD19 and CD3. Conclusion The combination of GM-CSF and IL-4 can promote the expansion and differentiation of DC in vitro.