贵州医科大学学报

2017, v.42;No.204(09) 993-997+1002

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Ⅰ型鼠尾胶原三维培养模型对树突状细胞形态及分泌能力的影响
The Effects of T-dimensional Rat Tail Collagen Ⅰ Cultivation Model on Morphology and Cytokine Secretion Capacity of Dendritic Cells

黄文竹;胡文慧;曾柱;
HUANG Wenzhu;HU Wenhui;ZENG Zhu;School of Biology and Engineering,Guizhou Medical University;Key Laboratory of Biology and Engineering,Guizhou Medical University;

摘要(Abstract):

目的:通过构建Ⅰ型鼠尾胶原三维(3D)培养模型,探索细胞外力学微环境对DCs产生的影响。方法:取健康人外周血来源的CD14+单核细胞,利用重组人白介素-4(rh IL-4)和重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)共同诱导5 d,获得未成熟DCs(im DCs),再利用重组人干扰素-γ(rh IFN-γ)和脂多糖(LPS)诱导im DCs为成熟DCs(mDCs);再利用鼠尾肌腱提取Ⅰ型鼠尾胶原体外培养DCs;FITC-Annexin-V和碘化丙啶(PI)标记细胞,检测DCs的凋亡率;酶联免疫吸附测定法检测DC的白细胞介素-12(IL-12)、IL-18和IL-1β的分泌水平。结果:与对照组相比,3D力学环境处理后细胞形态发生改变,细胞早期凋亡率降低,IL-18和IL-1β的分泌能力发生下调(P<0.05)。结论:细胞外3D力学微环境能够调控DCs的免疫功能。
Objective: To study the effects of extracellular mechanical microenvironment on dendritic cells( DCs) by establishing three-dimensional rat tail collagenⅠ cultivation model. Methods: CD14+peripheral blood mononuclear cells were induced to im DCs by recombinant human interleukin-4( rh IL-4) and recombinant human granulocyte-macrophage colony-stimulating factor( rh GM-CSF),and then induced to m DCs by recombinant human interferon γ( rh IFN-γ) and lipopolysaccharide( LPS). Rat tail collagenⅠwas extracted from rat tail tendon and then used to cultivate DCs in vitro. The cell apoptosis were measured by FITC-Annexin-V and PI labeled cells. The cytokine production levels were measured by Elisa kit. Results: Compared with control group,the 3D extracellular mechanical microenvironment could induce the morphological change,the ratio of early apoptosis were down-regulated,cytokine including IL-18 and IL-1β secretion capacity were down-regulated( P < 0. 05). Conclusions:The 3D extracellular mechanical microenvironment could regulate the immune function of DCs.

关键词(KeyWords): 树突状细胞;Ⅰ型鼠尾胶原;三维培养模型;凋亡;白细胞介素;免疫功能
dendritic cells;rat tail collagen Ⅰ;three-dimensional cultivation model;apoptosis;interleukin;immune function

Abstract:

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基金项目(Foundation): 国家自然科学基金(31260227,11162003,31771014,11762006);; 贵州省2011协同创新计划(2015-04);; 贵州省科技创新人才团队(2015-4021);; 教育部科学技术基金重点项目(210196);; 贵州省科技厅科技合作项目(LH-2016-7375);; 贵州医科大学博士启动基金(2014-024,2014-022)

作者(Author): 黄文竹;胡文慧;曾柱;
HUANG Wenzhu;HU Wenhui;ZENG Zhu;School of Biology and Engineering,Guizhou Medical University;Key Laboratory of Biology and Engineering,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2017.09.001

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