Luts法制备鹅骨髓源树突状细胞及其初步鉴定  

Preliminary Identification of Goose Bone Marrow-derived Dendritic Cells Prepared by Luts Method

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作  者:李睿婷 祝博森 郭子杰 朱婷 钟睿 洪伟鸣[1] 徐海 李玲[1] LI Ruiting;ZHU Bosen;GUO Zijie;ZHU Ting;ZHONG Rui;HONG Weiming;XU Hai;LI Ling(Jiangsu Agri-animal Husbandry Vocational College,Taizhou,Jiangsu 225300)

机构地区:[1]江苏农牧科技职业学院,江苏泰州225300

出  处:《中国家禽》2024年第4期54-59,共6页China Poultry

基  金:江苏农牧科技职业学院院级课题(NSF2023ZR13)。

摘  要:为制备鹅骨髓源树突状细胞(Dendritic cells,DCs),试验采用改良的Luts方法经粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素-4(IL-4)协同诱导骨髓单个核细胞分化成为DCs,通过形态观察、表面标志鉴定以及特征性生物功能分析初步鉴定培养获得的DCs。结果显示,培养至第10天细胞表面刺突、伪足明显,细胞集落生长,呈现典型树突状细胞形态;RT-qPCR检测到DCs成熟后表面标志物CD40、CD80和CD86转录水平的上调;培养至第8~10天细胞吞噬能力逐步达到峰值,至第12天细胞培养上清中IL-2和IFN-γ含量分别为(92.39±5.13)pg/mL和(1345.68±76.84)pg/mL;制备的DCs作为刺激细胞与淋巴细胞以1∶5混合培养时能显著刺激淋巴细胞增殖(P<0.05)。研究表明在体外成功诱导培养出鹅骨髓源DCs,制备的细胞表现出体内DCs所具备的部分生物学特性。To prepare goose bone marrow-derived dendritic cells(DCs),a modified Luts method was used to co-induce mononuclear bone marrow cells by granulocyte-macrophage colony-stimulating factor(GM-CSF)and interleukin-4(IL-4).Then the cultured DCs were identified by morphological observation,surface marker identification and characteristic biological func⁃tion analysis.The results showed that after 10 days of culture,a typical dendritic cell morphology was observed which manifested as obvious spike and pseudopodia on the cell surface and the cell colonies grew.The transcript levels of surface markers CD40,CD80 and CD86 were up-regulated after maturation of DCs by RT-qPCR detection.The phagocytic ability of the DCs gradually reached the peak after 8 to 10 days of cultivation.Meanwhile,the secrete IL-2 and IFN-γin the DCs culture supernatant on the 12th day reached(92.39±5.13)pg/mL and(1345.68±76.84)pg/mL,respectively.Additionally,a significantly proliferation of lym⁃phocytes was detected when co-cultured with the prepared DCs at a ratio of 1∶5(P<0.05).It was suggested that the goose bone marrow-derived dendritic cells were successfully introduction cultivated in vitro,and the prepared cells presented the typical bio⁃logical characteristic of in vivo DCs.

关 键 词:Luts法  树突状细胞 体外培养 鉴定 

分 类 号:S852.4[农业科学—基础兽医学]

 

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