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目的:探索骨髓间充质干细胞(BMSCs)及成骨诱导后的骨髓间充质干细胞(OM-BMSCs)对同种异体的外周血单个核细胞(PBMCs)的免疫调节作用。方法:分离比格犬BMSCs,成骨诱导培养基(ost eogenic medium,OM)进行成骨诱导,建立BMSCs、OM-BMSCs与同种异体PBMCs的共培养体系,分三种实验条件,共9组。A组:5×104受体PBMCs加入5×104供体PBMCs共培养;B组:A组组分加入已铺被1×104/孔BMSCs的96孔板;C组:A组组分加入已铺被1×104/孔OM-BMSCs的96孔板;D组:5×104受体PBMCs加入植物血凝素A(PHA)刺激物;E组、F组:将D组组分按B、C组方式处理;G组:5×104受体PBMCs加入IL-2炎症因子;H组、I组:G组组分按B、C组方式处理。A、D、G组分别为三个实验条件的对照组,单独接种1×104/孔BMSCs、OM-BMSCs作为空白对照组Bl ank1组、Bl ank2组。以上各组分别培养120h后,应用Cell Titer 96 Aqueous检测各组PBMCs的增殖情况。结果:在各实验条件下与相应对照组相比较,BMSCs与OM-BMSCs均能抑制双向混合淋巴细胞反应(MLR)中PBMCs的增殖,均能抑制经PHA刺激后PBMCs的增殖,均能抑制经IL-2刺激后PBMCs的增殖(P<0.01)。结论:BMSCs及成骨分化的BMSCs对刺激细胞、PHA、IL-2刺激的同种异体PBMSs的增殖均表现为抑制效应,成骨分化后的骨髓间充质干细胞对同种异体免疫反应仍具有免疫调节作用。
Objective: To explore the immunomodulatory effect of bone marrow mesenchymal stem cells (BMSCs) and osteogenic induced bone marrow mesenchymal stem cells (OM-BMSCs) on allogeneic peripheral blood mononuclear cells (PBMCs). Methods: BMSCs and osteogenic induction medium (OM) were isolated from Beagle dogs. Osteoblasts were induced to establish co-culture system of BMSCs and OM-BMSCs with allogeneic PBMCs. There were 9 experimental groups . Group A: 5 × 104 recipient PBMCs were added to 5 × 104 donor PBMCs for co-culture; Group B: Group A was added to 96-well plates plated with 1 × 104 BMSCs; Group C: Group A was added D group: 5 × 10 4 receptor PBMCs were added phytohemagglutinin A (PHA) stimulus; E group, F group: the group D components according to B, Group C: Group G: 5 × 104 recipients of PBMCs were added IL-2 inflammatory cytokines; Group H: Group I: Group G: Group B and Group C were treated. Groups A, D and G were control groups of three experimental conditions, respectively. Inoculated with 1 × 10 4 / well BMSCs and OM-BMSCs served as blank control group, Bl ank 1 group and Bl ank 2 group. After the above groups were cultured for 120h, Cell Titer 96 Aqueous was used to detect the proliferation of PBMCs in each group. Results: Compared with the corresponding control group, BMSCs and OM-BMSCs could inhibit the proliferation of PBMCs in the two-way mixed lymphocyte reaction (MLR) under all experimental conditions and both inhibited the proliferation of PBMCs stimulated by PHA, Proliferation of PBMCs after IL-2 stimulation (P <0.01). CONCLUSION: BMSCs and osteogenic differentiation of BMSCs can inhibit the proliferation of allogeneic PBMSs stimulated by PHA and IL-2. The osteogenic differentiation of BMSCs still has the effect of allogeneic immune response Immunomodulatory effect.