[1]刘 芳,乔艳艳,薛 洋,等.猪外周血单核细胞来源的树突状细胞体外诱导培养[J].江苏农业学报,2016,(03):588.[doi:10.3969/j.issn.1000-4440.2016.03.016]
　LIU Fang,QIAO Yan-yan,XUE Yang,et al.Porcine peripheral blood monocyte-derived dendritic cells cultured in vitro[J].,2016,(03):588.[doi:10.3969/j.issn.1000-4440.2016.03.016]
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猪外周血单核细胞来源的树突状细胞体外诱导培养()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年03期
页码:
588
栏目:
畜牧兽医·水产养殖
出版日期:
2016-06-30

文章信息/Info

Title:
Porcine peripheral blood monocyte-derived dendritic cells cultured in vitro
作者:
刘 芳1; 2;  乔艳艳1;  薛 洋1;  杜芳芳2;  张 俊1;  张红英1; 2;  王学兵1; 2
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
Author(s):
LIU Fang1; 2;  QIAO Yan-yan 1;  XUE Yang1;  DU Fang-fang2;  ZHANG Jun1;  ZHANG Hong-ying1; 2;  WANG Xue-bing1; 2
1.College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450000, China; 2.Key Laboratory for Animal-derived Food Safety of Henan Province, Zhengzhou 450000, China
关键词:
猪外周血单核细胞;  猪树突状细胞;  表型分子;  诱导培养
Keywords:
porcine peripheral blood monocyte;  dendritic cell;  phenotypic molecule;  induction
分类号:
S828; S852.4+3
DOI:
10.3969/j.issn.1000-4440.2016.03.016
文献标志码:
A
摘要:
为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,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(rpGM-CSF)和白细胞介素4(rpIL-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。结果表明,经体外诱导的细胞具有典型的树突状形态; 经脂多糖刺激的树突状细胞表型分子CD1a、CD80、CD86、SLAII、CD172a与未经刺激的树突状细胞有明显增高,其吞噬能力有所下降,刺激同种异体T淋巴细胞的能力增强。本研究成功建立了猪MoDC的体外诱导培养方法,为进一步研究其在机体免疫调节和抗病毒感染中的作用奠定了基础。
Abstract:
To develop a method for maturation induction of dendritic cells(DCs) in vitro, porcine peripheral blood monocyte-derived DCs(MoDCs)were induced in the presence of rpGM-CSF, rpIL4 and lipopolysaccharide(LPS). The cells were harvested to detect the morphological, phenotypic and functional characteristics. FITC-dextran endocytic activity and the surface molecules of MoDCs were measured with flow cytometer, and T lymphocyte proliferation following MoDCs stimulation were detected by CCK-8 assay. The induced MoDCs showed typical dendritic morphology, and the expressions of the surface molecules CD1a, CD80, CD86, SLAII, and CD172a were increased by LPS treatment from 53.8%,42.4%,23.4%,54.2%,48.2% to 55.2%,53.6%,43.6%,65.4%,61.1%, respectively. The phagocytic ability of non-LPS-treated DC to uptake FITC-dextran was higher than that of LPS-treated DC. The proliferation of allogenetic T lymphocyte was enhanced in LPS-treated DCs. In conclusion, the porcine MoDC was successfully induced and cultured in vitro, laying a foundation for further study on the role of porcine MoDC in immune regulation and anti-virus infection.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2015-10-09
基金项目:河南省基础与前沿技术研究项目(152300410076); 河南省自然科学基金研究项目(132300410114); 河南省科技攻关计划(152102110048)
作者简介::刘 芳(1979-),女,河南邓州人,博士,主要从事中药免疫调节作用研究。(E-mail)girlliufang@163.com。乔艳艳为共同第一作者。
通讯作者:王学兵,(E-mail)xbwang74@163.com
更新日期/Last Update: 2016-06-30