大鼠骨髓间充质干细胞与Sertoli细胞体外共培育的免疫负调作用

Immunosuppressive effects of rat bone marrow mesenchymal stem cells in the co-culture system with Sertoli cells in vitro

目的探讨骨髓间充质干细胞(BMSCs)与Sertoli细胞(SCs)体外共培育时对免疫应答的调节作用,为两者联合应用于移植修复提供线索。方法用密度梯度离心法分离SD大鼠脾脏单个核细胞(简称L),羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记细胞后,将其加入按照不同的比例混合的BMSCs与SCs共培育体系,设立L细胞为对照组,L+刀豆蛋白A(ConA)、BMSCs+L+ConA、SCs+L+ConA、BMSCs+SCs+L+ConA四大组细胞为实验组。培养4 d,通过流式细胞术分析T淋巴细胞的增殖情况。结果单个核细胞培养体系中加入ConA可以显著刺激T淋巴细胞增殖。BMSCs、SCs均可抑制淋巴细胞增殖,抑制作用随加入BMSCs和SCs的比例增加而增强,BMSCs和SCs混合培养后对淋巴细胞增殖抑制作用进一步增强,且在某些浓度时呈现协同性。结论 BMSCs与SCs体外共培育时可增强免疫抑制作用。

Objective To explore the regulating for immune response by bone marrow mesenchymal stem cells BMSCs and Sertoli cells co-culture system in vitro,providing clues to the transplantation of the combination of the two methods. Methods Splen mononuclear cells of normal SD rat were isolated using ficoll-hypaque density-gradient centrifugation method,labeled with carboxy-fluorescein diacetate succinimidyl ester（ CFSE） and then added to bone marrow mesenchymal stem cells（ BMSCs） and Sertoli cells（ SCs） co-cultivation system in different proportions. One control group（ L）,as well as four treatment groups（ L ＋ ConA,BMSCs ＋ L ＋ ConA,SCs ＋ L ＋ ConA,BMSCs ＋ SCs ＋ L ＋ ConA） were set up for experimental study. Cells were cultured for 4 days. Flow cytometry was adopted to evaluate the proliferation of T lymphocytes. Results BMSCs or SCs showed inhibition of lymphocyte proliferation induced by ConA in a dose-dependent manner. The inhibition of lymphocyte proliferation could be further enhanced in BMSCs and SCs co-culture system. Conclusions BMSCs and SCs co-culture in vitro was able to enhance the suppression of T cell immune response.