Culture of Porcine Peripheral Blood Monocyte-derived Dendritic Cells in vitro  

猪外周血单核细胞来源的树突状细胞体外诱导培养（英文）

作　　者：乔艳艳[1] 薛洋[1] 闫延华[1] 杜芳芳[2] 张俊[1] 张红英[1,2] 王学兵[1,2] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450000 [2]河南省动物性食品安全重点实验室,河南郑州450000

出　　处：《Agricultural Science & Technology》2016年第10期2338-2342,共5页农业科学与技术（英文版）

基　　金：Supported by Fundamental and Advanced Research Projects of Henan Province(152300410076,2015-2017);Key Science and Technology Program of Henan Province(152102110048,2015-2017)~~

摘　　要：[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.[目的]建立猪外周血单核细胞来源的树突状细胞(MoDC)体外培养模型。[方法]从猪外周新鲜血液中分离外周血单核细胞(PBMC),通过贴壁法获得树突状细胞前体细胞,采用重组猪的集落共刺激因子(rp GM-SCF)和重组猪的白细胞介素4(rp IL-4)双因子诱导及脂多糖(LPS)刺激成熟法,收集不同时间段的细胞,利用扫描电子显微镜观察其形态,流式细胞分析仪检测表面分子表达率及其对FITC-dextran的吞噬能力,混合淋巴细胞反应检测细胞对同种异体T细胞的刺激能力。[结果]经体外诱导的DC具有典型的树突状形态;LPS刺激的DC表型分子CD1a、CD80、CD86、SLAII、CD172a与LPS未刺激的DC有明显增高,其吞噬能力有所下降,刺激同种异体T淋巴细胞的能力有所增强。[结论]该试验成功建立了猪MoDC的体外诱导培养方法,为进一步研究猪MoDC在机体免疫调节和抗病毒感染中的作用奠定了基础。

关 键 词：Porcine peripheral blood monocyte INDUCTION Porcine dendritic cell Phenotypic molecule 

分 类 号：S828[农业科学—畜牧学]