小鼠骨髓源树突状细胞的体外培养及鉴定

目的建立小鼠骨髓源树突状细胞（bonetnarrowdendriticcell，BMDC）的培养方法并对其表型和功能进行鉴定．方法无菌取BALB/c小鼠股骨、胫骨中的骨髓细胞，以粒一巨噬细胞集落刺激因子体外诱导分化为BMDC，倒置显微镜动态观察BMDC增殖和形态变化情况，流式细胞术分析细胞表型，并检测其抗原吞噬功能．结果小鼠骨髓细胞体外诱导可获得大量未成熟和成熟BMDC，呈现典型的树突状形态．未成熟BMDC的细胞表型为CDllchighCD40lowcD86lowMHC—Illow，具有较强的抗原吞噬能力．未成熟BMDC经细菌脂多糖刺激后可分化为高表达CD11c、CD40、CD86及MHC—II类分子的成熟BMDC．结论体外诱导培养可获得小鼠骨髓来源的未成熟和成熟DC。

Cultivation and Identification of Dendritic Cells from Mouse Bone Marrow in Vitro

Objective To establish a method of cultivation of dendritic cells （DC） from mouse bone marrow in vitro and identify their phenotype and function. Methods Under aseptic condition, bone marrow cells were extracted from the tibia and femur bones of BALB/c mice. Bone marrow cells were cultured with recombinant mouse granulocyte-maerophage colony-stimulating factor （rmGM-CSF） in vitro. The expansion and morphological changes of DC were observed with light inverted microscope. Phenotype was identified with flow cytometry and biological function was studied with antigen phagoeytosis test. Results A large number of immature and mature DC with typical dendritic morphological characteristics could be generated from murine bone marrow. Immature DC, which had high expression in CD1 le and low expression in CD40, MHC-II and CD86, could phagoeytize antigen. Mature DC, which could be induced from immature DC by lipopolysaecharides, had high expression in CD1 lc, CD40, CD86 and MHC-II molecules. Conclusion Immature and mature DC can be generated from mouse bone marrow cells through eytokine induction in vitro and be used for further study associated with DC.