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

摘要

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(rhIL-4)和重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)共同诱导5d,获得未成熟DCs(imDCs),再利用重组人干扰素-γ(rhIFN-γ)和脂多糖(LPS)诱导imDCs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 collagen Ⅰ cultivation model.Methods:CD14 + peripheral blood mononuclear cells were induced to imDCs by recombinant human interleukin-4(rhIL-4) and recombinant human granulocyte-macrophage colony-stimulating factor(rhGM-CSF),and then induced to mDCs by recombinant human interferon γ(rhIFN-γ) and lipopolysaccharide (LPS).Rat tail collagen Ⅰ was extracted from rat tail tendon and then used to cultivate DCs in vitro.The cell apop-tosis 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 3 D 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.