不同培养基对人外周血单核细胞来源树突状细胞发育的影响

摘要

This study was aimed to investigate the effect of RPMI 1640 and IMDM on the development of human peripheral blood monocyte-derived dendritic cells.Under the same cytokines and culture conditions, the different medium types were tested, and the morphology of mature and immature dendritic cells was observed by microscopy, the cell phenotype and endocytosis ability were detected by flow cytometry.Furthermore, the immunoregulatory function of various DC was analyzed by mixed lymphocyte reaction (MLR), the expression of cytokine in culture supernatant of MLR system was also analyzed by Bio-plex technology.The results showed that there were no difference in morphology, CD14, CD83 expression and endocytosis ability between MDM-culmred DC and RPMI-1640 medium-cultured DC, but there was a lower expression of CD1 a in IMDM-cultured DC.Moreover, DC cultured with IMDM displayed a significant reduction in stimulating T cell proliferation, and highly expressed IL-6, IL-8 and IL-10, but low expressed IL-12.It is concluded that the different cultural mediums can induce DC with different functions and DC cultured with IMDM may correlated with induction of immune tolerance.The results of this study will provide a new idea for DC clinical application.%本研究旨在探讨RPMI 1640和IMDM两种培养液对诱导人外周血单核细胞向树突状细胞(DC)发育分化的影响.利用GM-CSF+IL-4细胞因子组合,在培养条件一致的前提下,改变培养液种类,通过对成熟、未成熟DC形态观察,用流式细胞术检测细胞表型和吞噬能力、应用混合琳巴细胞反应(MLR)检测其剌激T细胞的增殖能力,用悬浮芯片技术检测DC与同种异体T细胞共培养后上清中所含细胞因子的变化,分析不同培养液对以DC功能的影响.研究结果表明,两种培养液培养所得的DC在形态上无显著差异,DC的吞噬能力以及CD14和CD83的表达亦无显著差异,但应用IMDM培养的DC的CDa表达明显降低,且对T细胞增殖的刺激能力也显著降低;IMDM培养的DC高表达IL-6、IL-8和IL-10,而IL-12的表达则显著降低.结论:不同的培养体系可获得功能不同的DC,IMDM培养的DC可能与免疫耐受有关,本研究结果为DC在临床上的应用提供了新思路.