论文部分内容阅读
目的:建立一种简便高效的体外扩增培养小鼠骨髓树突状细胞(bone marrow dendritic cell,BMDC)的方法,为树突状细胞(dendritic cells,DCs)的理论研究与临床应用提供实验工具。方法:联合应用重组小鼠粒细胞-巨噬细胞集落刺激因子(rmGM-CSF)、重组小鼠白介素(rmIL-4)诱导培养小鼠骨髓单个核细胞,终末加入重组小鼠肿瘤坏死因子(rmTNF-α),从形态学表型及功能方面进行检测。结果:培养3天后可见大量贴壁细胞及细胞集落形成,第5天,可见典型的树突状突起,光镜下动态观察树突状细胞DC的形态变化,流式细胞仪检测各组DC加入TNF-α前后表面标志物CD11c、CD86的表达,加入TNF-α后CD86的阳性表达率明显增高,CD11c的阳性表达率变化无统计学意义。结论:此方法能在体外诱导和扩增出大量骨髓源性DC,为抗肿瘤疫苗研究及临床应用奠定基础。
OBJECTIVE: To establish a simple and efficient method for culturing mouse bone marrow dendritic cells (BMDC) in vitro and to provide experimental tools for the theoretical research and clinical application of dendritic cells (DCs) . Methods: The mouse bone marrow mononuclear cells were induced by recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin-4 (IL-4), and the recombinant mouse tumor necrosis factor rmTNF-α), from morphological phenotype and function testing. RESULTS: A large number of adherent cells and colony formation were observed after 3 days of culture. On the fifth day, typical dendritic processes were observed. The morphological changes of dendritic cells were observed under light microscope. DCs of DCs were detected by flow cytometry The expression of CD11c and CD86 on the surface of tumor necrosis factor-α (TNF-α) before and after TNF-α expression was significantly increased after adding TNF-α, while the positive expression rate of CD11c had no statistical significance. Conclusion: This method can induce and amplify a large number of bone marrow-derived DCs in vitro and lay a foundation for the study of antitumor vaccine and clinical application.