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目的 探讨由动员人外周造血祖细胞体外培养扩增获得的树突状细胞（DC）的生物学特性, 为临床应用肿瘤树突状细胞疫苗建立制备方法。方法 用CS-3000血细胞分离机分离预经干细胞动员的患者外周血单个核细胞（PBMC），去除淋巴细胞和单核细胞，阴性选择得到外周造血祖细胞，加入rhGM-CSF和rhIL-4培养10~14 d成为树突状细胞，然后进行形态学检测、细胞表型和T细胞刺激能力分析。结果 由外周造血祖细胞来源的树突状细胞可以达到(0.5~1.0)×108数量级；高倍镜和电镜照片显示典型的树突状细胞形态和超微结构；流式细胞仪分析显示细胞高表达HLA-Ⅰ、Ⅱ类抗原和共刺激分子CD80、CD86；同种混合淋巴细胞反应显示细胞具有高效的刺激静止T淋巴细胞活化增殖的能力。结论 用血细胞分离机分离动员外周造血祖细胞体外定向诱导制备树突状细胞可以作为制备肿瘤疫苗的新途径。
Objective To investigate the biological characteristics of dendritic cells (DCs) obtained by mobilizing peripheral hematopoietic progenitors in vitro and to establish methods for clinical application of tumor dendritic cell vaccines. METHODS: Peripheral blood mononuclear cells (PBMCs) were removed from peripheral blood mononuclear cells (PBMCs) pretreated with stem cells by a CS-3000 blood cell separator. Lymphocytes and monocytes were removed. Peripheral hematopoietic progenitor cells were selected by negative selection and cultured with rhGM-CSF and rhIL-4. Dendritic cells became ~14 days old, followed by morphological examination, cell phenotype, and T cell stimulating ability analysis. Results Dendritic cells derived from peripheral hematopoietic progenitor cells could reach the order of (0.5~1.0) x 108; high densitometer and electron micrographs showed typical dendritic cell morphology and ultrastructure; flow cytometric analysis showed high expression of cells. HLA-I, II antigens and co-stimulatory molecules CD80, CD86; the same mixed lymphocyte reaction showed that cells have the ability to stimulate the activation and proliferation of resting T lymphocytes. Conclusion The in vitro induction of dendritic cells by mobilizing peripheral hematopoietic progenitor cells from blood cell separators can be used as a new way to prepare tumor vaccines.