背景:胰岛移植为有望根治糖尿病的一种方法,而移植排斥问题阻碍了移植的开展,诱导受体对供体移植物的免疫耐受是解决同种异体移植排斥反应的关键。目的:观察骨髓间充质干细胞输注对糖尿病小型猪同种异体胰岛移植物存活时间的影响。方法:无菌条件下抽取动物骨髓,离心弃上清。取单核细胞层,洗涤后加入L-DMEM培养基,调整细胞浓度为1×10^9L^-1。培养48h后换液去除未贴壁的细胞,依据贴壁培养法分离和扩增间充质干细胞。取第5代细胞作为移植供源,调整细胞浓度为1×10^10L^-1。四氧嘧啶制备糖尿病小型猪模型,随机分为骨髓细胞组和干细胞+骨髓细胞组,经肝门静脉分别植入胰岛细胞、骨髓细胞,胰岛细胞、骨髓细胞和骨髓间充质干细胞。结果与结论:骨髓间充质干细胞联合骨髓细胞输注较单纯骨髓细胞输注能显著延长同种异体小型猪胰岛移植物存活时间(P〈0.05),同时受体血清胰岛素能较长时间维持较高水平。提示骨髓间充质干细胞输注能显著延长猪胰岛移植物存活时间,并促进胰岛移植物正常功能的发挥。
BACKGROUND: Islet transplantation is a promising method used to radically cure diabetes mellitus. The immune rejection caused by transplantation hinders the development of transplantation. The immune tolerance of recipients to donor grafts is a key to solving the problem of allotransplantation rejection reaction. OBJECTIVE: To investigate the influence of infusion of bone marrow mesenchymal stem cells on the survival time of islet allograft in the minipigs after islet allotransplantation. METHODS: Under sterile condition, minipig bone marrow was taken and the supernatant was discarded after centrifugation. After washed with L-DMEM culture medium, mononuclear cells were adjusted to be 1 ×10^9L^-1. After 48-hour culture, nonadherent cells were discarded and mesenchymal stem cells were isolated and amplified. Passage 5 cells were taken as donors and cell concentration was adjusted to be1 ×10^10L^-1. Minipig models of alloxan-induced diabetes mellitus were prepared and then randomly divided into two groups: bone marrow cells and bone marrow cells + mesenchymal stem cells. In the bone marrow cells group, islet cells and bone marrow cells were infused. In the bone marrow cells + mesenchymal stem cells group, islet cells, bone marrow cells and bone marrow mesenchymal stem cells were infused. RESULTS AND CONCLUSION: Compared with infusion of simple bone marrow cells, infusion of bone marrow mesenchymal stem cells and bone marrow ceils significantly prolonged the survival time of islet allografts (P 〈 0.05) and maintained high serum insulin level for a longer time in diabetic minipigs. These findings suggest that bone marrow mesenchymal stem cell infusion can significantly prolong the survival time of islet allografts and contribute to the normal function of islet allografts in diabetic minipigs.