人外周血中树突状细胞的诱导及鉴定

Generation of Dendritic Cells from Healthy Human Peripheral Blood

目的 从健康人外周血中诱导高纯度树突状细胞 (dendritic cell,DC)。方法 用贴壁法从健康人外周血获取单核细胞 ,对传统的诱导方法加以改进 ,经 GM- CSF(10 0 ng/ ml)和 IL- 4(5 0 0 U/ m l)持续诱导 7天 ,在此期间不再补充新的细胞因子、新鲜培养基和促成熟因子 ,以获取高纯度 DC。结果 经上述方法处理后 ,从健康人外周血中诱导出了大量高纯度 DC,其能高表达 HL A- 、 类分子、共刺激分子和粘附分子 ,显示出成熟 DC的特征。这些 DC能强烈诱导同种异体淋巴细胞的增埴 ,其内吞能力在第 3天达最高 ,之后明显下降。结论 本研究所建立的从外周血获取大量成熟 DC的方法经济、简便 ,为

Objective To get high quality and sufficient numbers of mature dendritic cells (DC) from healthy human peripheral blood. Methods Cultured plastic adherent monocytes were isolated from healthy human peripheral blood by use of granulocyte monocyte clony stimulating factor and Interleukin 4 for 7 days without fed with fresh medium and cytokine. Results A large number of DCs with high purity were generated. These DCs expressed HLA Ⅰ,Ⅱmolecules, Costimulating molecules and adherent molecules highly, showing the characteristics of mature DC. These DCs could stimulate proliferation of allogenic T lymphocytes and had active endocytosis ability which peaked at the third day in culture but decreased afterwards. Conclusion These results provide evidence that CD monocytes isolated from peripheral blood monocytes exhibit dendritic cells characteristics and may facilitate further studies of DC and its clinical application.