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目的    研究不同年龄阶段口腔癌患者外周血单核细胞来源的树突状细胞（dendritic cell，DC）的形态、表型差异并分析其临床意义。方法    将2007—2010年兰州大学第一医院及第二医院口腔科住院的口腔鳞癌初诊患者30例，分为40～50岁及≥60岁2组，每组各15例。取患者外周血单核细胞经重组人粒细胞集落刺激因子（rhGM-CSF）、重组人白细胞介素4（rhIL-4）及肿瘤坏死因子-α（TNF-α）诱导培养，获取成熟树突状细胞，光镜观察细胞形态，流式细胞仪检测细胞表面协同刺激分子CD1a、CD83、CD80、CD86的表达。结果    培养7d后，2组均诱导出典型形态和表型的树突状细胞，其中≥60岁组DC培养过程中细胞逐渐死亡，培养至第7天大部分细胞死亡，获得的DC数量较少，有3例未能培养出DC；而40～50岁组培养细胞的活力保持较好，至第7天均可获得数量较多的DC。结论    2组口腔鳞癌患者外周血单核细胞来源的DC经细胞因子诱导培养，均能扩增出成熟DC，但较40～50岁组，≥60岁组外周血培养的DC体外生长活力明显降低，可能与老年患者免疫功能明显降低有关。

In vitro culture and phenotype analysis of monocyte-derived dendritic cell from peripheral blood of patients with oral carcinoma of different age groups

Objective To investigate the phenotype of induced dendritic cell （DC） derived from peripheral blood of patients with oral carcinoma of different age groups. Methods Totally 30 patients with oral carcinoma at stage Ⅲor above were assigned to study group（≥60 years old,n = 15 ;40～50 years old,n = 15）. Leukocytes were isolated by density centrifugation. Adherent fractions were incubated in present of rhGM-CSF,rhIL-4 and TNF-α for 7 days in vitro. The morphological change of the cultured cells was observed by light microscopy. The phenotype expressions of CD83,CD1a,CD86,CD80 were analysized by flow cytometry. Results Typical DCs were obtained from two groups after 7 days culture. In the group of ≥60years,most of the DCs were nonviable at the 7th day after culture,and no DCs were detected in 3 subjects. On the other hand,in the group of 40～50 years,the DCs grew prosperous in all the subjects. At the 7th day after culture,DCs could be seen in every subject. Conclusion DC derived from the two groups of oral cancer patient＇ s peripheral blood monocyte might be matured,but the growth velocity,vigour and survival time of the DCs are lower in the group of ≥60 years than that in the middle-aged,which may be related to the immunological function impairment in the very elderly.