摘要目的 建立荧光示踪骨髓重建裸小鼠模型及其在胶质瘤干祖细胞招募骨髓源细胞研究中的应用.方法 不同放射剂量辐照裸小鼠并收集辐照后外周血及骨髓数据确定清髓性骨髓毁损的最佳剂量;辐照后24 h内经尾静脉移植表达绿色荧光蛋白(GFP)的裸小鼠骨髓细胞.流式检测GFP+细胞在骨髓及外周血中的分布,比较移植前后骨髓细胞集落形成能力、骨髓切片来评价重建骨髓功能;在此模型上原位接种红色荧光蛋白(RFP+)的胶质瘤干祖细胞(SU3-RFP)观察其对骨髓源细胞的影响.结果 6 Gy辐照后3～5d裸小鼠外周血白细胞数接近于0;骨髓细胞集落数与正常组比较显著减少.骨髓移植后裸小鼠外周血白细胞在28 d恢复到辐照前水平;流式结果显示骨髓和外周血中GFP+细胞分别为(90.95±2.05)％和(83.62±0.73)％;骨髓细胞集落数与正常组比较差异无统计学意义(P＞0.05).SU3-RFP原位移植瘤中发现GFP+骨髓源细胞特异性迁徙到肿瘤区域.结论 6 Gy是裸小鼠清髓性骨髓毁损的最佳剂量;GFP+骨髓细胞裸小鼠模型可用于示踪研究肿瘤与宿主骨髓源细胞的关系;胶质瘤干祖细胞对骨髓源细胞具有特异性招募作用.

abstractsObjective The nude mice model with green fluorescent protein (GFP) + bone marrow was established and used to indicate host myeloid cells recruited by glioma stem/progenitor cells.Methods Different doses X-ray were used to irradiate the nude mice.The data of peripheral blood and bone marrow were detected to determine the optimal dose destroying myeloablative bone marrow function of nude mice.Allogeneic transplantation with GFP + bone marrow cells (BMCs) were executed through the caudal vein within 24 h.The flow cytometry was applied to detect the distribution of GFP + cells in bone marrow and peripheral blood.And the bone marrow function was evaluated by the comparison of the forming ability of bone marrow colony,leukocyte in peripheral blood and the pathological section of bone marrow before and after transplantation.Glioma stem/progenitor cells expressed red fluorescent protein (RFP) were orthotopic transplanted into nude mice model with GFP + bone marrow.Their effect on bone marrow-oriented cells was disclosed by fluorescence microscope.Results The leukocyte in peripheral blood declined to 0 in 3 to 5 days after 6 Gy irradiation,and the quantity of bone marrow cell colony reduced significantly compared with normal group.The leukocyte in peripheral blood recovered normally within 28 days after GFP + bone marrow transplantion.The proportion of GFP + cells in bone marrow and peripheral blood were (90.95 ± 2.05) ％ and (83.62 ± 0.73)％ respectively.The quantity of bone marrow cell colony has no statistical difference with normal group.GFP + bone marrow-oriented cells migrated specifically to glioma region.Conclusion 6 Gy is the optimal dose for establishment of the bone marrow damage reconstituted nude mice model.GFP + bone marrow nude mice can be used to reveal the effects of tumors on host bone marrow cells.Host bone marrow-oriented cells were recruited specially by glioma stem/progenitor cells.