小鼠骨髓间充质干细胞免疫学特性及其对急性移植物抗宿主病的影响

发布时间：2018-10-17 12:25

【摘要】：目的:分离、培养小鼠骨髓间充质干细胞(mMSCs);体外验证mMSCs抑制有丝分裂原和异基因抗原诱发的脾细胞免疫反应性增殖;动物实验证明mMSCs促进植入,减轻急性移植物抗宿主病(GVHD)。方法:1.全骨髓贴壁法和单克隆培养法分离、纯化mMSCs,体外连续传代,鉴定其生物学特性。2.MTT法检测mMSCs对脾细胞增殖的抑制作用;对细胞亚群及细胞因子分泌的影响。3.观察mMSCs对急性GVHD临床表现、组织病理的影响;动态监测其对T细胞亚群及细胞因子分泌的影响。结果:1.成功获得mMSCs,体外连续传代30代以上仍能保持生物学特性稳定,具有多向分化潜能,无致瘤性。2.mMSCs抑制脾细胞的免疫反应性增殖,使CD4~+/CD8~+比值升高、CD4~+CD25~+细胞比例增加。3.动物实验中,1×10~5mMSCs共移植可以促进异基因造血植入,减轻GVHD。5×10~6mMSCs单一输注可促进受致死剂量照射的异基因小鼠自体造血恢复,无GVHD表现。结论:mMSCs参与移植可以促进植入及造血恢复,减轻GVHD。
[Abstract]:Objective: to isolate and culture mouse bone marrow mesenchymal stem cells (mMSCs);) and verify in vitro that mMSCs inhibits the proliferation of splenocytes immunoreactive induced by mitogen and allogeneic antigens. Animal experiments have shown that mMSCs promotes implantation and alleviates (GVHD). Of acute graft-versus-host disease. Methods: 1. Whole bone marrow adherent method and monoclonal culture method were used to isolate and purify mMSCs, in vitro to identify its biological characteristics. 2.MTT assay was used to detect the inhibitory effect of mMSCs on spleen cell proliferation and the effect of mMSCs on cell subsets and cytokine secretion. 3. To observe the effect of mMSCs on the clinical manifestation, histopathology and T cell subsets and cytokine secretion of acute GVHD. The result is 1: 1. The successful passage of mMSCs, in vitro for more than 30 passages could maintain stable biological characteristics, have multidirectional differentiation potential, and have no tumorigenesis. 2.mMSCs inhibited the immune reactive proliferation of splenocytes, increased the ratio of CD4~ / CD8~, and increased the proportion of CD4~ CD25~ cells. In animal experiment, 1 脳 10~5mMSCs co-transplantation could promote allogeneic hematopoietic implantation, and alleviate GVHD.5 脳 10~6mMSCs single infusion could promote autologous hematopoiesis recovery of allogeneic mice exposed to lethal dose, without GVHD manifestation. Conclusion: mMSCs involved in transplantation can promote implantation and hematopoietic recovery, and alleviate GVHD..
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R392

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