小鼠骨髓腔内输注人脐血造血干／祖细胞植活水平的观察  

The observation of engraftment of human umbilical cord blood-derived hematopoietic stem/progenitor cells in xenotransplanted NOD/SCID mouse model by intra-bone marrow injection

作　　者：高静韬[1] 卢士红[1] 李妍涵[1] 杨舟[1] 许静[1] 郑以州[1] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所血液病医院实验血液学国家重点实验室,天津300020

出　　处：《中华血液学杂志》2008年第6期361-365,共5页Chinese Journal of Hematology

基　　金：基金项目：国家自然科学基金（30571762）

摘　　要：目的探讨小鼠骨髓腔内输注能否增强人脐血造血干／祖细胞（HS／PC）异种移植的植活能力。方法将不同数量（1×10^3、1×10^4、0.5×10^5、1×10^5、5×10^5）的人脐血CD34^＋细胞经尾静脉和骨髓腔途径移植入经亚致死剂量照射的NOD／SCID小鼠。于指定时间点处死小鼠，用PCR法检测人17号染色体α-微卫星特异性片段及用流式细胞术检测人源CD45^＋细胞，观察脐血CD34^＋细胞在移植小鼠左、右两侧胫骨和股骨及脾脏等部位的归巢及其长期植活能力。结果异种移植后24h，经骨髓腔移植5×10^5 CD34^＋细胞的小鼠肝、脾、肺组织，外周血，左右两侧胫骨和股骨、骨髓细胞均表达人17号染色体α-卫星特异性片段。不同数量的人脐血CD34^＋细胞经骨髓腔途径移植入NOD／SCID小鼠后，8周时其植活良好（检测部位包括输注部位右侧胫骨，以及非输注部位右侧股骨、左侧胫骨、左侧股骨、脾脏及外周血）。分别经尾静脉和骨髓腔两种途径输注同一来源的人脐血CD34^＋细胞1.0×10^5，8周时两组间人造血细胞植活水平分别为（44．063±20．095）％和（45．881±22．316）％，差异无统计学意义（P〈0．05）；而将移植CD34^＋细胞数降至1.0×10^4时，则经骨髓腔途径输注的人脐血CD34^＋细胞小鼠植活水平（54．019±31．338）％显著优于尾静脉输注途径[（12．197±10．350）％，P〈0．01）]；当CD34^＋细胞输注量降至1.0×10^3时，仅有骨髓腔内输注组小鼠能见到人脐血CD34^＋细胞植活，且植活部位通常为非输注部位骨骼。结论小鼠骨髓腔内移植能够增强人脐血造血干／祖细胞的植活水平。Objective To explore whether intra-bone marrow injection strategy could promote the en-graftment of human umbilical cord blood derived hematopoietic stem/progenitor cells （HS/PC） in xenotrans-planted NOD/SCID mouse model. Methods Aliquots containing 1×10^3、1×10^4、0.5×10^5、1×10^5、and 5×10^5 human umbilical cord blood （hUCB） CD34^＋ cells were transplanted into sublethally irradiated NOD/ SCID mice via intra-venous（IV） and intra-bone marrow（iBM） injection. The homing and long-term engraft- ment capabilities of hUCB CD34^＋ cells from right tibia, right femur , left tibia, left femur and spleen were detected by PCR 24h after xenotransplantation and by FACS 8-week after xenotransplantation. Results Tissues of liver, spleen ,lungs, or cells from peripheral blood, right tibia, right femur, left tibia and left femur 24 hours after xenotransplantation in iBM injecting 5×10^5 CD34^＋ cells recipients expressed human chromo- some 17 specific or-satellite fragment. 8-week engraftment of human cells was observed and engraftment level indicated dose-dependent effect in injected bone （right tibia） as well as non-injected bones （including right femur, left tibia and left femur）, spleen and peripheral blood in all iBM recipients. 8-week engraftment levels of human cells were （44.063±20.095） % and （45.881±22.316） % for IV and iBM groups respectively, when transplanted with 1×10^5 hUCB CD34^＋ cells, being no statistical difference （ P 〉 0.05 ）. More superior 8-week engraftment levels of human cells were observed in iBM recipients [（54. 019±31.338）%] than in Ⅳ recipients [（12.197±10.350）%] when transplanted with 1.0×10^ CD34^ cells （ P 〈 0.01 ）. Human cell engraftment was observed in iBM but not in IV recipients when transplanted with 1.0×10^ CD34^ cells, and was usually obsened in non-injected bones. Conclusion Intra-bone marrow strategy can efficiently increase the engraftment of umbilical cord blood derived hematopoietic stern/progenitor cells in xen

关 键 词：造血干细胞移植 骨髓腔注射 小鼠 NOD/SCID 造血重建 

分 类 号：R686[医药卫生—骨科学]