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目的探讨间充质干细胞株C3H10T1/2对小鼠骨髓来源树突状细胞(dendritic cells,DCs)体外分化、成熟的影响。方法采用密度梯度离心法分离小鼠骨髓细胞,在mGM-CSF和mIL-4刺激下获得大量未成熟DCs。将DCs和C3H10T1/2细胞共培养2d,同时加入LPS刺激,分别通过倒置显微镜、流式细胞仪,混合淋巴反应以及ELISA检测DCs的成熟。结果经C3H10T1/2细胞体外共培养的DCs,形态观察呈散在分布,细胞边缘圆滑,流式细胞仪检测显示其低表达CD11c、MHC-Ⅱ、CD80、CD86、CD40;共培养组的DCs刺激同种异型小鼠脾细胞增殖的能力在各个浓度均明显低于对照组(P<0.05或0.01);ELISA检测培养上清中IFN-γ和IL-10的浓度也明显低于对照组(P<0.05或0.01)。结论间充质干细胞株C3H10T1/2可以明显抑制小鼠骨髓来源DCs的体外成熟。
Objective To investigate the effect of mesenchymal stem cell line C3H10T1 / 2 on the differentiation and maturation of murine bone marrow-derived dendritic cells (DCs) in vitro. Methods Murine bone marrow cells were isolated by density gradient centrifugation, and a large number of immature DCs were obtained under stimulation of mGM-CSF and mIL-4. DCs and C3H10T1 / 2 cells were co-cultured for 2 days and stimulated with LPS. The maturation of DCs was detected by inverted microscope, flow cytometry, mixed lymphocyte reaction and ELISA, respectively. Results DCs co-cultured with C3H10T1 / 2 cells showed scattered distribution with smooth edges and showed low expression of CD11c, MHC-Ⅱ, CD80, CD86 and CD40 by flow cytometry The ability of proliferation of allogeneic mouse spleen cells was significantly lower than that of the control group (P <0.05 or 0.01). The concentrations of IFN-γ and IL-10 in the culture supernatants were also significantly lower than those of the control group (P < 0.05 or 0.01). Conclusion Mesenchymal stem cell line C3H10T1 / 2 can significantly inhibit the maturation of murine bone marrow-derived DCs in vitro.