小鼠骨髓树突状细胞体外培养扩增与生物学特性研究

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（bonemarrowdendriticcell，BMDC）的方法，为树突状细胞（dendl’iticcells，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一巨噬细胞集落刺激网子（rmGM—CSF）、蘑纽小鼠白介素（rmlL-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的衷达，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。

Amplification in Vitro and Identification of Biological Characteristics of Dendritic Cell from Mouse Bone Marrow

Objective： To establish an eflhctive anti convenient method of inducing and culturing dendritic cells from mouse bone marrow in vitro, and provide a theoretical basis for the theoretical study and clinical application of dendritic cells. Methods： Mononuclear cells isolated torm mouse bone marrow were induced into dendritic, cells by being cultured with rmGM-CSF and rmIL-4, rmTNF-α was added at last. Morphology, pbenotype and function of induced DCs were then examined. Results： A large number of colonies and anchorage-dependent cells were formed 3 days after in vitro culturing. Typical morphology with dendritic processes could be observed at Ihe fifth day of culturing. Then the rnovphology changes of DC were observed under light microscope. CD11c ,CD86 expression levels before and after addition of TNF-α were com- pared . The positive rates of CD86 after adding TNF-α were significantly higher than the level before adding of TNF-α. No obvious ebangc was observed of Ihe positive tales of CDIIe. before and after the adding of TNF-α. Conclusion： A large number of DCs can be induced and amplifild from murine bone marrow in vitro by this way. It can lay a foundation for anti- tumor vacine research resaeach and clinical appilication.