异基因骨髓间充质干细胞逆向嵌合体诱导小鼠皮肤移植耐受研究  

Allogenic BMSCs Reverse Chimerism Inducing the Tolerance of Xeno-skin Graft in Mouse Model

作　　者：龙丹[1] 吴文桥[1] 卢俊[1] 冯莉[1] 李幼平[1] 李胜富[1] 

机构地区：[1]四川大学华西医院卫生部移植工程与移植免疫重点实验室,成都610041

出　　处：《四川大学学报（医学版）》2013年第2期165-169,共5页Journal of Sichuan University(Medical Sciences)

基　　金：教育部-博士点基金(No.20060610060)资助

摘　　要：目的探讨骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)诱导移植免疫耐受的可行性,建立小鼠异种逆向嵌合皮肤移植模型。方法以GFP-C57BL/10雄鼠为BMSCs供者,选择免疫缺陷型BALB/C-nu/nu雌鼠为BMSCs受者,移植BMSCs,建立骨髓嵌合模型;通过RT-PCR检测Y染色体性别决定区基因(SRY)、免疫组化检测GFP蛋白表达量,确定嵌合产生的最佳时间。实验共分3组:将接受BMSCs移植一定时间后的BALB/C-nu/nu雌鼠的皮肤移植给C57BL/10雄鼠(实验组),将未接受BMSCs移植的BALB/C-nu/nu雌鼠的皮肤移植给C57BL/10雄鼠(排斥组),C57BL/10小鼠间进行同种异体皮肤移植(对照组)。观察移植皮肤存活情况,检测存活皮肤的血管形成情况,以此探讨逆向嵌合皮肤移植的可行性。结果成功建立骨髓嵌合模型,用1×106 BMSCs移植后4周,SRY基因和GFP蛋白的表达量最高,分别为1.22±0.10、458.0±3.4,与移植后1周、2周和6周比较差异均有统计学意义(P<0.05),说明嵌合产生的最佳时间为移植后4周。实验组小鼠供者来源皮肤移植物存活>14d,排斥组平均为5d。结论 BMSCs移植后可形成嵌合体,并可逆向嵌合移植诱导异种小鼠皮肤移植耐受。Objective To investigate the feasibility of bone marrow mesenchymal stem cells （BMSCs） in inducing immune tolerance and to establish the mouse model of reverse chimerism in xeno-skin transplantation. Methods The mouse model of bone marrow-chimerism was established with immuncompromised BAI.B/C-nu/nu female mice by receiving the transplantation of BMSCs from green fluorescent protein （GFP）-C57BL/10 male mice, the optimized chimeric time was identified by RT-PCR testing of SRY gene and immunohistochemistry measurement of GFP expression. In the experiment group, GFP-C57BL/10 male mice received the transplantation of the skin from immuncompromised BALB/C-nu/nu female mice with BMSCs bone marrow chimerism. In the rejection group, GFP-C57BL/10 male mice received the transplantation of the skin from immuncompromised BALB/C-nu/nu female mice without BMSCs bone marrow-chimerism. In the control group, allo-transplantation of skin was performed in GFP-C57BL/10 male mice. Histological study was performed to investigate the survival rate and angiogenesis of the transplanted skin. Results The bone marrow chimeric model was established, the expressions of SRY gene and GFP protein reached the highest level at four weeks （1.22 ± 0. 10 458.0± 3.4） post-transplanted with BMSCs （106）, which was significantly different in comparison with those at one week, two weeks and six weeks post- transplantation（P〈0.05）. Four-weeks after transplantation was further confirmed as the optimized chimeric time. The mean survival time of donor skin graft 〉 14 d in the experimental group, while it only was 5 d in the rejection group. Conclusion The bone marrow-chimerism can be formed in the recipient by donor BMSCs transplantation, which can further induce tolerance of mice xeno-skin transplantation by reverse chimerism.

关 键 词：GFP小鼠 骨髓 干细胞移植 嵌合体 皮肤移植耐受 

分 类 号：R392[医药卫生—免疫学]