中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 小鼠骨髓源CD103~+ DC分离培养及LPS对其形态与功能特征的影响 浙江农业学报 2018 (7) 1122-1131
Position: Home > Articles > In vitro culture of CD103~+ DCs from mouse bone marrow and effects of LPS on its morphology and functional characteristics Acta Agriculturae Zhejiangensis 2018 (7) 1122-1131

小鼠骨髓源CD103~+ DC分离培养及LPS对其形态与功能特征的影响

作  者：
侯艳华;张凯;王磊;孙静;王旭荣;张康;王学智;李建喜;张景艳
关键词：
C57BL/6小鼠;骨髓源CD103+DC;分离培养;LPS;功能特征
摘  要：
旨在建立C57BL/6小鼠骨髓源CD103~+树突状细胞(CD103~+dendritic cell,CD103~+DC)分离培养方法,阐述LPS对其形态与功能特征的影响。在无菌条件下分离C57BL/6小鼠骨髓细胞,并用重组GM-CSF和FLT3L对其进行体外联合诱导培养;利用光镜、扫描电镜、荧光显微镜和流式细胞术,分别对LPS作用前后细胞形态、表型及功能进行了分析。结果表明,细胞培养至第3天有零星集落出现,第13天后集落开始分散,可见典型的树突状突起,第15天后可得到大量的CD103~+DC,加LPS刺激培养24 h后细胞表面树突样结构更加明显;分离培养的骨髓细胞能够表达表面分子CD103,其表达率达90%以上。RPMI-1640组(LPS未刺激组)可吞噬VOA的CD103~+DC比例为25.70%,能够表达MHC-Ⅱ和CD83阳性细胞分别为41.31%和13.79%;LPS刺激组可吞噬VOA的CD103~+DC比例为10.33%,能够表达MHC-Ⅱ和CD83的阳性细胞分别为68.10%和24.71%。MTT法检测结果显示,经LPS处理的CD103~+DC刺激T细胞增殖的能力明显增强。综上所述,分离于C57BL/6小鼠的骨髓细胞,可在体外经FLT3L和GM-CSF共同诱导培养出CD103~+DC,LPS可促进CD103~+DC的成熟。
译  名：
In vitro culture of CD103~+ DCs from mouse bone marrow and effects of LPS on its morphology and functional characteristics
作  者：
HOU Yanhua;ZHANG Kai;WANG Lei;SUN Jing;WANG Xurong;ZHANG Kang;WANG Xuezhi;LI Jianxi;ZHANG Jingyan;Lanzhou Institute of Husbandry and Pharmaceutical Science of CAAS,Center of Engineering and Technology of TCVM in Gansu Province;
单  位：
HOU Yanhua%ZHANG Kai%WANG Lei%SUN Jing%WANG Xurong%ZHANG Kang%WANG Xuezhi%LI Jianxi%ZHANG Jingyan%Lanzhou Institute of Husbandry and Pharmaceutical Science of CAAS,Center of Engineering and Technology of TCVM in Gansu Province
关键词：
C57BL/6 mice;;bone marrow derived CD103+ DC;;isolation and culture;;LPS;;functional characteristics
摘  要：
In order to establish a method of inducing and culturing CD103~+dendritic cells( CD103~+DCs) from C57 BL/6 mice bone marrow in vitro,and investigate the effects of LPS on their morphology and functional feature.The bone marrow cells( BMCs) were isolated and cultured in the presence of recombinant granulocyte-macrophage colony-stimulating factor( GM-CSF) and fms-like tyrosine kinase 3 ligand( FLT3 L),and then observed the effect of LPS on their morphology,phenotype and function. The results showed that the modality of BMCs diversified after being cultured in GM-CSF and FLT3 L for 3 d,and grew clustered-liked. As time went on,the cell colonies became lar-ger and larger. When culturing for 13 d,the BMC colonies started separating,typical morphology with dendritic processes could be observed. A number of morphologically typical dendritic cells were observed after culturing for 15 d. The dendritic structure became more obvious after BMCs were treated with LPS for 24 h under scan electron microscope. The induced BMCs were able to express CD103 on the cell surface,and the positive rate of CD103~+DC was over 90%. The percentage of phagocytosis of CD103~+DCs was 25. 70% in the group of RPMI-1640,but the percentage of phagocytosis in the group of LPS dropped to 10. 33%. In the group of RPMI-1640,41. 31% CD103~+DCs expressed MHC-II,13. 79% expressed CD83. The CD103~+DCs treated by LPS showed a higher MHC-II expression rate of 68. 10%,and increase expression rate in CD83( 24. 71%). MTT assay showed that the ability of CD103~+DC treated with LPS to stimulate the proliferation of the naive allergenic T cells was stronger than those CD103~+DC untreated with LPS. In conclusion,a large number of dendritic cells can be generated by culturing BMCs from C57 BL/6 mice in vitro,and LPS shows positive effect on CD103~+DC mature.

相似文章

导出引用
计量
文章访问数: 6
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46