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目的:探讨从人外周血单核细胞体外培养扩增的树突状细胞(dendriticcell,DC)形态学特征及其活性。方法:人外周血常规分离单个核细胞,粘附6h后去除悬浮细胞,以细胞因子粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)进行诱导培养7d,并进行形态学特征,细胞表型和淋巴细胞刺激能力鉴定。结果:培养1周即可得到大量DC,形态学观察可见细胞形态不规则,细胞表面大量突起,为典型的DC特征,免疫组化和间接免疫荧光显示CD1a阳性表达率达80%～95%,并能刺激同种淋巴细胞增殖反应。结论:人外周血单核细胞可经GM-CSF、IL-4诱导培养成DC,具有典型的树突状细胞的形态学特征及抗原呈递能力。
Objective: To investigate the morphological characteristics and activity of dendritic cells (DCs) cultured in vitro from human peripheral blood mononuclear cells. Methods: Mononuclear cells were isolated from human peripheral blood. After 6 h of adhesion, suspended cells were removed and induced by GM-CSF and IL-4 7d, and morphological features, cell phenotype and lymphocyte stimulating ability identification. Results: A large number of DCs could be obtained after one week of culture. Morphological observation showed irregular cell morphology and massive cell surface protuberances, which were typical DC characteristics. Immunohistochemistry and indirect immunofluorescence showed that CD1a positive rate was 80% -95% And can stimulate all kinds of lymphocyte proliferation reaction. CONCLUSION: Human peripheral blood mononuclear cells can be cultured into DCs induced by GM-CSF and IL-4, with the typical morphological characteristics and antigen presenting ability of dendritic cells.