同种异基因小鼠骨髓腔内脐血移植模型的建立及其对造血干细胞植入的影响  

Establishment of a Murine Model for Allogeneic Umbilical Cord Blood Transplantation by Intra-bone Marrow Injection and Its Effects on the Engraftment of HSCs

作　　者：蔡耘[1] 黄绍良[1] 陈凤英[2] 黄文革[2] 魏菁[3] 车丽洪[4] 

机构地区：[1]中山大学附属第二医院儿科 [2]中山大学实验动物中心 [3]中山大学附属第二医院医学研究中心 [4]中山大学病理教研室,广东广州510120

出　　处：《中山大学学报（医学科学版）》2005年第6期644-650,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：卫生部临床学科重点基金(2004468)

摘　　要：【目的】建立同种异基因小鼠骨髓腔内脐血移植模型，研究该技术对造血干细胞(HSC)归巢、植入率、移植后免疫造血重建、移植物抗宿主病(GVHD)等方面的影响。【方法】用C57BL／6胎鼠及新生鼠外周血(FNPB)作供体，以单侧胫骨骨髓腔内注射(IBMI)和尾静脉注射(IV)两种途径移植经亚致死量^(60)Coγ射线辐照预处理的BALB／c鼠。200只受鼠随机分为4组：骨髓腔内注射组(IBM组)；尾静脉注射组(IV组)；放疗对照组；正常对照组，每组50只。用组织冰冻切片和流式细胞术动态了解CFSE标记FNPB在受体内的分布变化，并观察移植后受鼠存活状况、植入水平、造血与免疫功能恢复及GVHD情况。【结果】①荧光际记FNPB体内动力学显示直至输注后72h，供体FNPB经IBMI后主要积聚于注射侧骨髓腔内，肺部滞留很少。而IV组及IBM组非注射侧骨髓中的FNPB细胞数显著少于IBM组注射侧，IV组非造血组织器官如肺部有明显供体细胞滞留。②同种异基因小鼠CBT模型中IBM组造血、免疫功能的恢复明显快于IV组，无GVHD，移植后90d存活率达到90％，注射侧胫骨植入水平(29.53±6.64)％，较IV组有显著改善。③IBM输注侧骨髓中供体HSCs植入水平及造血重建速度明显优于非注射侧骨髓。【结论】成功建立同种异基因小鼠骨髓腔内脐血移植模型，并证实IBMI途径对促进HSCs植入，造血免疫功能重建，提高UCBT效果方面优于IV途径，多部位IBMI可能更有利于HSC归巢骨髓。[Objective] To establish a murine model for allogeneic umbilical cord blood transplantation by intra-bone marrow injection, investigate its effects on the homing and engraftment of HSCs, hematopoietic and immunological reconstruction and GVHD of recipient mice after transplantation. [Methods] BALB/C recipient mice conditioned with sublethal dose ^60Co γ-ray radiotherapy were transplanted with FNPB from C57BL/6 mice by intrabone marrow injection or via the tail vein. Two hundred recipient mice were divided into four groups at random： intrabone marrow injection group （IBM group）, intravenous injection group （Ⅳ group）, irradiated control group, normal group. There were 50 recipient mice in every group. The distribution of CFSE-labeled FNPB inside the recipients was observed in frozen sections of different organs or by flow cytometry. The survival rate, engraftment level, recovery of hematopoietic and immunological function and GVHD of recipient mice were studied. [Results] （1） Tracing of the CFSE-labeled FNPB showed that by 72 hours after infusion the IBM-injected FNPB mainly accumulated in the BM cavity of the injected side tibia and few was trapped in the lung, but the amount of FNPB inside the noninjected side tibia or the tibia of Ⅳ group was significantly less than that of the injected side tibia. Lots of FNPB were obviously sequestrated by other nonhematopoietic organs like the lungs by Ⅳ injection. （2） The recovery of hematopoietic and immunological function of IBM group was remarkably faster than Ⅳ group and its engraftment level and survival rate were improved without GVHD. By ＋90 d, the survival rate was 90% and the level of H-2Db＋ cells in the injected side tibia BMNCs of IBM group was（29.53±6.64）%. （3）The engraftment level of HSCs and the hematopoietic reconstruction of the IBM-injected side tibia were much better than the noninjected side. [Conclusion] The murine model for allogeneic umbilical cord blood transplantation by intra-bone marrow injection was establish

关 键 词：骨髓 脐血移植 动物模型 造血干细胞 

分 类 号：R457[医药卫生—治疗学]