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

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 dendriticcells （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 micro- environment 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.