Culture and identification of chicken bone marrow-derived dendritic cells in vitro | 鸡骨髓源树突状细胞体外诱导培养及鉴定

Chinese. 为建立鸡骨髓源树突状细胞（DC，dendriticceils）体外培养方法，用重组鸡粒细胞一巨噬细胞集落刺激因子（rGM―CSF）和重组鸡白细胞介素4（rIL-4）体外诱导鸡骨髓细胞分化为DC，然后通过形态观察、表型鉴定及功能分析来初步鉴定所培养的DC。试验结果表明：培养7d后，光学冠微镜下观察到细胞表而不规则，有显著的树突状突起，呈典型的DC形态，流式细胞仪测得细胞表面CD11c和MHCⅡ分子的表达量分别为69．3％和63．0％。经脂多糖或CpG―ODN刺激24h后的DC，其表面成熟分子标志C1M0和CD86上调表达，同时其刺激同种异体T细胞增殖的能力显著增强（P〈0．01）。结论：本试验建立的方法能在体外制备出大量具有较高纯度的鸡骨髓源DC，并具有体内DC的生物学特秆。

English. To create a method for cultivation of chicken bone marrow-derived dendritic （：ells （ DC ） in vitro, chicken bone marrow cells were isolated and cultured in the medium with recombinant chicken interleukin（rIL-4） and granulocyte-macrophage colony-stimulating factor（rGM-CSF） in vitro. Then the DC was preliminarily identified by morphologic ,phenotypic and functional assays in vitro. Results： after 7 days of culture, under the inverted microscope,there were many dendrite-like processes on the surface of the cuhured （;ells and the cells displayed the typical morphology of DC. The expressions of CDI lc and MHC Ⅱ on the surface of the cuhured cell were 69.3% and 63.0%. The CD40 and CD86 expressions on the surface of DC which were stimulated by lipopolysaccharide（LPS） or CpG-ODN for 24 hours were up-regulated, and the DC stimulated by LPS or CpG-ODN could significantly induce the proliferation of allogenic mixed T lymphocytes. Conclusion：a large quantity of chicken bone marrow-derived DC with higher purity could be obtained in vitro by this way used in our study, and the cultured cells display the typical biological characteristics of DC in vivo.