一种大鼠带血管的骨髓移植模型

A vascularized bone marrow transplantation model in rat

目的构建一种简单可靠的带血管的骨髓移植模型。方法将供体带血管蒂的股骨通过显微血管吻合的方式移植到受体腹股沟区，20只Lewis近交系大鼠随机分3组：同基因移植组Lewis→Lewis；排斥组Lewis→BN；免疫抑制组Lewis→BN，术后给予环孢素A。通过大体和病理学检查观察各组移植物存活情况，流式细胞计数监测外周血嵌合水平。结果术后30d，同基因移植组股骨存活良好，骨髓苏木素．伊红（HE）染色与正常骨髓无异，而排斥组第7天即发生显著的排斥反应，骨髓细胞明显减少、坏死，外周血嵌合水平几乎为0；和同系移植组一样，免疫抑制组术后30d移植物存活良好，术后1、2、3、4周外周血中可检测到供体特异性嵌合，嵌合水平分别为（4．7±2．0）％、（2．2±1．2）％、（1．8±0．9）％、（1．5±0．3）％。结论带血管蒂的股骨移植是一种简便可靠的骨髓移植模型，可作为一种新的骨髓移植方法诱导免疫耐受。

Objective To establish a simple and reliable vascularized bone marrow transplantation （VBMT） model as a new method for bone marrow transplantation. Methods The femoral bones were transplanted into the recipient inguinal regions and their nutrient vessels were anastomosed with recipients vessels. Twenty Lewis rats were divided into 3 groups. In syngeneic group, femoral bone transplantation was performed between Lewis rats. In both rejection and immunosuppression groups, BN rats were used as donors and Lewis rats as recipients. Rats in rejection group received no treatment, and those in immuno- suppression group were given cyclosporine A after allotransplantation. The viability of the all grafts was e- valuated grossly and morphologically, and the chimerism was determined by using flow cytometry. Results The bone grafts in syngeneic group remained viable and the bone marrow maintained normal morphologically at 30th day post-transplant, but the grafts in rejection group were rejected at 7th day after transplan- tation, and mmber of bone marrow cells was decreased sharpiy almost no chimerism. In accord with syngeneic grafts, the grafts in immunosuppression group showed normal appearance of the viable tis- sue. Donor-specific chimerism was confirmed in peripheral blood at 1st, 2nd, 3rd, and 4th week post- transplant, and the ehimerism level was detected at （4. 7 ± 2. 0） %, （2. 2 ± 1.2 ） %, （ 1.8 ±0.9 ） % and （ 1.5 ± 0. 3 ） % respectively. Conclusion Vascularized femoral bone grafting is a simple and reliable method for the transplantation of bone marrow, and may provide a useful tool for the study of immune toler- ance.