第三方骨髓间充质干细胞对同种异体皮肤移植的影响

目的 探讨第三方骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)对同种异体皮肤移植的影响.方法 40只雌性C57BL/6和50只雄性BALB/C小鼠分别作为皮肤移植的供体和受体.将50只BALB/C小鼠随机分为5组,每组10只:空白对照组,直接进行皮肤移植;给予受体环磷酰胺组(Cyelophosphamide,CP组);给予受体SD大鼠BMSCs移植组(SD-BMSCs组);给予受体CP+SD-BMSCs移植组(CP+SD-BMSCs组);CM-DiI荧光标记组.CP+SD-BMSCs组给予大剂量CP腹腔注射,200 mg/kg,2 d,移植当天自受体小鼠尾静脉注射1×10~6个SD-BMSCs.检测项目包括:观察皮片存活时间、单向混合淋巴反应、组织HE染色、组织荧光镜检,流式细胞仪检测受体外周血单核细胞中SD-BMSCs的含量等.结果 CP+SD-BMSCs组皮肤移植物存活时间(14.5±1.9)d,空白对照组为(8.O±0.8)d,CP组为(11.1±1.4)d,SD-BMSCs组为(10.9±1.4)d,CP+SD-BMSCs组皮肤移植物存活时间明显比后3组延长(P<0.05).混合淋巴反应结果显示,BMSCs对淋巴细胞的增殖抑制作用存在量效关系.HE染色发现CP+SD-BMSCs组不但有血管生成,淋巴细胞浸润也较少.CM-DiI标记的SD-BMSCs能够在BALB/c小鼠体内存活30 d以上,SD-BMSCs组与CP+SD-BMSCs组外周血流式细胞仪均检测到SD-BMSCs.结论 BMSCs能够在异种受体体内存活较长时间;第三方BMSCs能够延长同种异体移植物的存活时间;BMSCs能够抑制异种T淋巴细胞的活化增殖.

Impact of third-party bone marrow mesenchymal stem cells on allogenic skin transplantation

Objective To investigate the effect of the third-party bone marrow-derived mesenchymal stem cells (BMSCs) on the allogeneic skin transplantation. Methods 40 female C57BL/ 6 mice and 50 male BALB/ C mice were respectively used as donors and recipients of skin transplantation. SO BALB/ C mice were divided randomly into 5 groups : Blank control group , Cyclophosphamide group , BMSCs group , Cyclophosphamide + BMSCs group and CM-DiI staining group , with 10 mice in each group. Before skin transplantation, high-dose abdominal injection of Cyclophosphamide ( 200 mg/kg,2 d) was performed in recipient mice. On the transplantation day, a bonus of 1 × 10~6 BMSCs of the SD rat (SD-BMSCs) were injected through the tail vein. The observation of skin grafts, mixed lymphocyte culture ( MLC) , HE staining,the observation of CM-Dil-labeled SD-BMSCs and FACS were used. Results The skin graft survival time was significantly prolonged in the Cyclophosphamide + BMSCs group, as compared with the blank control group, the Cyclophosphamide group, the BMSCs group respectively. When BMSC and lymphocyte mixed at the ratio of 1:1 and 1:10, rat BMSCs inhibited T lymphocyte proliferation. More angiogenesis and less lymphocyte infiltration were found in the experimental group than them in other groups. Red fluorescent cells were found in CM-DiI staining group under long-term observation. The SD-BMSCs can be detected by flow cytometry in the cell group and the Cyclophosphamide + BMSCs group. Conclusions BMSCs can survive in the heterogeneic recipient body; the third-party BMSCs transplantation can prolong skin graft survival time; BMSCs can inhibit T lymphocyte activation and proliferation.