成人外周血来源树突状细胞体外诱导培养动态监测

Culture of Dendritic Cells in Adult Peripheral Blood In Vitro

目的:分析体外诱导培养树突状细胞(dendritic cells,DCs)表面分子及成熟度的动态变化;方法:利用流式细胞仪(fluores-cence activated cell sorter,FACS)检测分析DCs表面成熟标志分子CD83和T细胞辅助分子CD58、CD54、CD40、CD80、CD86、HLA-DR、HLA-ABC表达水平;结果:体外诱导培养5 d细胞即高表达DCs标志分子CD11c,细胞成熟度与细胞表面T细胞辅助分子的表达水平随培养天数的增加有一定提高,但显著低于大肠杆菌LPS刺激的DCs;结论:诱导培养6 d DCs表型为CD83low、CD58low、CD54 low、CD40 low、CD80 low、CD86 low、HLA-DRhigh、HLA-ABChigh,为不成熟DCs。

Objective：To analyse the mature dynamic and phenotype molecular changes of human peripheral blood-derived den-dritic cells（DCs） in vitro;Methods：Monocytes,which were isolated from human peripheral blood mononuclear cells,were cultured in medium containing IL-4 and GM-CSF.After 5～6 days of culture to detects the mature marker CD83 and co-stimulatory molecules：CD58,CD54,CD80,CD86,CD40,HLA-DR,HLA-ABC,which was determined by fluorescence activated cell sorter（FACS）;Results：After 5～6 days of culture,more than 95% of cells converted into iDCs with phenotype of CD11c＋,CD54low,CD83low,CD40low,CD58low,CD86low,and CD80low,which was determined by fluorescence activated cell sorter（FACS）.E.coli LPS-pulsed DCs exhibit-ed significantly high expression of mature marker CD83 and co-stimulatory molecules：CD58,CD54,CD80,CD86,CD40.Conclusions：These results suggest that human monocyte-derived DCs are immature after 5～6 days of culture.