鸡骨髓源树突状细胞的诱导分化及鉴定

The development of chicken bone marrow-derived dendritic cells inducing with cytokines in vitro

为建立鸡骨髓源树突状细胞(Dcs)的体外培养和鉴定方法,本研究无菌抽取10日龄健康SPF雏鸡的骨髓,体外分离纯化骨髓细胞,将其培养于含有重组的人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和白细胞介素4(rhIL4)的RPMI 1640营养液中,诱导培养鸡骨髓源DCs。采用显微镜观察其体外培养过程中的形态特征及流式细胞仪(FACS)分析其表型特征。结果显示:鸡骨髓细胞经诱导培养,由单一的形态结构转变成多形态结构,培养至8 d后分化成具有典型形态的DCs。FACS分析发现诱导细胞表达了DCs的分子标记CD11c,显示成熟DCs的特征。本研究通过体外培养鸡骨髓细胞成功诱导分化成DCs,为进一步研究鸡DCs在家禽病原致免疫抑制机理中的作用提供了研究材料。

To explore the methods of inducing dendritic cells（DCs） from chicken bone marrow cells in vitro,chicken bone marrow cells were taken from 10-day-old chicken and cultured in the presence of recombinant human granulocyte-macrophage factor（rhGM-CSF） and rhIL-4.The results demonstrated that the cells showed typical morphologic and phenotypic characteristics of DCs after 8 day＇s cultivation and expressed higher level of CD11c as the surface marker of DCs detected by FACS staining with PE-conjugated anti-CD11c antibody.The feasibility for inducing chicken bone marrow cells with GM-CSF and IL-4 to generate DCs was demonstrated in this study.Thus,GM-CSF and IL-4 are required for the differentiation of the bone marrow-derived DCs in chicken.