不同来源人树突状细胞的培养诱导及鉴定

目的通过对不同来源的人树突状细胞的体外培养与诱导,建立树突状细胞的稳定体系,为更进一步实验打好基础。方法分离人外周血单个核细胞(PBMC),用不同树突状细胞亚群的特异性表面标记抗体磁珠分离树突状细胞前体,通过加入不同的刺激分子,分别诱导未成熟、成熟及活化等不同阶段的树突状细胞。结果与未成熟树突状细胞相比,成熟活化型树突状细胞表达HLA-DR、CD86以及CD40等共刺激分子明显升高,其中MoDC对成熟标记物CD83的表达水平亦较高。此外,两者在形态上也有明显的差异。结论通过对不同亚群树突状细胞的体外诱导、培养和鉴定,建立了较为成熟的研究树突状细胞的平台,为深入开展工作打下基础。

Isolation, culture, induction and identification of different subsets of human dendritic cells

Objective To isolate, culture and induce different subsets of dendritic cells from human peripheral blood mononuclear cells (PBMC). Methods PBMCs were prepared from buffy coats of peripheral blood by centrifugation through a Ficoll density gradient. Then dendritic cells were isolated using the specific monoclonal antibody magnetic microbeads. After adding different cytokines, cells were differentiated into different stages. Results The purity of dendritic cells was greater than 95% as assessed by flow cytometry. Mature denendritic cells expressed higher levels of HLA-DR, CD86, CD40 than immature dendritic cells. There were also morphological differences between two subsets of dendritic cells. Conclusion We have developed the methodology for isolating, culturing and inducing the dendritic cells of different stages in vitro.