小鼠骨髓间充质干细胞输注抑制同种异体皮片移植排斥反应的实验研究

Experimental study on suppression of skin allograft rejection by bone marrow mesenchymal stem cells (BMSCs) infusion in mice

  • 摘要:
    背景 异体皮片移植是大面积烧伤早期的重要治疗方法,骨髓间充质干细胞可降低移植排斥反应,有望为烧伤患者异体皮片排斥反应的治疗提供新的思路。目的 探究对异体皮片移植的受体小鼠输注供体来源的骨髓间充质干细胞,抑制同种异体皮片移植排斥反应的可行性。方法 体外分离培养小鼠骨髓间充质干细胞,通过流式细胞术验证其表面标志物CD34、CD45、CD105和CD106的表达情况,通过CCK8法和CTV法检测骨髓间充质干细胞抑制T淋巴细胞增殖的能力。C57BL/6J小鼠作为受体鼠,随机分为空白对照组、同基因移植组、对照组 (异体移植)、BMSCs组 (异体移植+BMSCs),观察皮片存活时间,检测淋巴结和脾脏内调节性T细胞的比例变化,以验证骨髓间充质干细胞抑制同种异体移植排斥反应的能力。结果 本研究通过原代培养获得的骨髓间充质干细胞,低表达CD45和CD34,高表达CD105和CD106,符合骨髓间充质干细胞的标志物表达。在T细胞增殖抑制培养体系中,骨髓间充质干细胞能够有效抑制T细胞增殖,且骨髓间充质干细胞比例越高,对T细胞增殖的抑制作用就越强。预先给予骨髓间充质干细胞注射的小鼠,皮片存活时间长于单纯异体皮片移植的小鼠。注射骨髓间充质干细胞的移植受体小鼠,术后淋巴结和脾脏中调节性T细胞的比例较单纯皮片移植组小鼠更高。结论 本研究成功分离并培养小鼠骨髓间充质干细胞,通过体外实验证明骨髓间充质干细胞可抑制T细胞增殖,动物实验证实骨髓间充质干细胞可延长异体皮片存活时间,并提高移植受体淋巴结和脾脏中调节性T细胞的比例。

     

    Abstract: Background Allogeneic skin graft is an important treatment in the early stage of large area burn. Bone marrow mesenchymal stem cells (BMSCs) can reduce transplant rejection and may provide new therapeutic ideas for the treatment of allogeneic skin graft rejection in burn patients. Objective To investigate the feasibility of infusing donor-derived BMSCs into recipient mice with skin allograft transplantation to inhibit allograft rejection.Methods Mouse BMSCs were isolated and cultured in vitro to verify the expression of surface markers. The ability of BMSCs to inhibit the proliferation of T lymphocytes was detected by CCK8 and CTV assays. C57BL/6J mice were randomly divided into Blank group, Autograft group, Control group (allograft) and BMSCs group (allograft +BMSCs) to verify the ability of BMSCs to inhibit allogeneic allograft rejection. The survival time of skin grafts was observed, and the proportion of regulatory T cells in lymph nodes and spleens was detected. Results In this study, BMSCs obtained by primary culture showed low expression of CD45 and CD34, and high expression of CD105 and CD106, which was consistent with the marker expression of BMSCs. In the T cell proliferation suppression culture system, mesenchymal stem cells can effectively suppressive T cell proliferation, and the higher the proportion of mesenchymal stem cells, the stronger the inhibitory effect on T cell proliferation. Mice injected with mesenchymal stem cells showed longer skin graft survival than mice transplanted with allogeneic skin grafts alone. Skin allograft recipients injected with mesenchymal stem cells showed higher regulatory T cells proportion in the lymph nodes and spleens after surgery than mice in the skin graft alone group.Conclusion BMSCs have been successfully isolated and cultured in this study, the results demonstrated that BMSCs could effectively inhibit the proliferation of T lymphocytes in vitro, prolong the survival time of grafts, and increase the proportion of regulatory T cells in lymph nodes and spleens after infusion into mice.

     

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