人外周血来源树突状细胞诱导培养及鉴定的实验研究

Studying on the Induction and Identification of Dendritic Cells from Human Peripheral Blood

目的探讨树突状细胞(DC)体外培养的方法。方法用重组人粒细胞巨噬细胞集落刺激因子、重组人白细胞介素4、重组人肿瘤坏死因子α等细胞因子自外周血单个核细胞诱导分离DC,流式细胞仪检测其表面标志,四氮唑蓝法测定其刺激同种异体淋巴细胞增殖能力。结果体外培养第10天,大量细胞出现典型的树突状形态;流式细胞仪检测高表达CD80、CD86、CD83、CD40、CD1a、人白细胞DR抗原;在体外能强烈刺激同种异体混合淋巴细胞增殖反应。结论该方法可获得较高纯度典型的DC,为DC的临床免疫治疗提供实验依据。

Objective To explore the cultivated methods of dendritic cells in vitro.Methods DC were isolated from human peripheral blood mononuclear cells by recombinant human gametocyte/macrophage colony stimulating factor for injection,recombinant human interleukin 4 and recombinant human tumor necrosis factorα.The surface signal of DC was analysed by fluorescence-actived cell sorting,and the abilities to stimulate the proliferation of allogenic lymphocyte by DC were evaluated by MTT assay.Results A number of cells had typical morphology characteristics on the 10th day.The cells expressed high level of CD80,CD86,CD83,CD40,CD1a,HLA-DR and were potent to stimulate the proliferation of allogenic lymphocytes.Conclusion This method obtained the highly effective and typical DC,and can be used for clinical immunotherapy.