摘要
为了探讨鸡骨髓源树突状细胞(bone marrow-derived dendritic cells,BMDCs)的体外诱导培养方法,并分析鸡树突状细胞(DCs)的主要生物学特性,将用淋巴细胞分离液分离的鸡骨髓细胞经差速贴壁纯化后获得单个核细胞,然后经粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)体外诱导分化,细胞培养后第7天再加入脂多糖(LPS)继续培养48 h刺激成熟,分别于倒置显微镜和电子显微镜下进行形态变化观察,同时应用流式细胞仪对鸡骨髓源树突状细胞表面标志进行分析、鉴定。结果显示鸡骨髓细胞培养7 d后,细胞体积增大,细胞表面长出树突状小突起,细胞表面高表达MHCⅡ和CD11c分子,经LPS刺激后,突起伸长变粗,扫描电镜观察呈典型的树突状细胞形态,细胞表面MHCⅡ表达显著升高,依据上述方法成功获得大量而高纯度的鸡骨髓源树突状细胞,为进一步研究禽类DCs的生物学功能及其在某些疾病发生中的作用奠定了基础。
In order to develop the method of induction and cultivation chicken bone marrow-derived dendritic cells(BMDCs)in vitro,and analyze the main characteristics of DCs,the bone marrow precursors were collected by differential adherence method after separating the chicken bone marrow cells using Lymphoprep.Then induction and differentiation of the bone marrow precursors were conducted by using granulocyte macrophage colony stimulating factor(GM-CSF)and interleukin-4(IL-4).On day 7 of culture,the cells were stimulated with lipopolysaccharide(LPS)for 48 h,then the DCs were identified by morphologic and phenotypic characteristics using inverted phase-contrast microscope,scanning electron microscope and flow cytometry.The results showed that the cells volume enlarged,and the surface of the cells were dendritically shaped,phenotypic analyses of non-stimulated cells showed high expression of MHC classⅡand CD11c.After stimulating by LPS,the veils became larger,these cells appeared a typical morphological feature of BMDCs under electron microscope,and the expression of MHC classⅡwas significantly increased.These data indicated that we succesfully obtained a large quantity of BMDCs with higher purity according to this method.This study laid a foundation for further research on the biological function of BMDCs in poultry and the role of BMDCs in the occurrence of certain diseases.
作者
张淑君
周放
吴乐
高雪丽
刘超男
吕晓萍
郑世民
ZHANG Shujun;ZHOU Fang;WU Le;GAO Xueli;LIU Chaonan;L Xiaoping;ZHENG Shimin(Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2019年第12期2571-2577,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31472169)
关键词
鸡
骨髓源树突状细胞
鉴定
生物学特征
chicken
bone marrow-derived dendritic cells
identification
biological characteristics
作者简介
张淑君(1993-),女,内蒙古乌兰察布人,硕士生,主要从事动物免疫病研究工作,E-mail:1059098614@qq.com;通信作者:郑世民,主要从事免疫病理学研究,E-mail:zhengshiminbl@sohu.com。
