小鼠骨髓腔内输注人脐血造血干/祖细胞植活水平的观察

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(1×103、1×104、0.5×105、1×105、5×105)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5×105 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1.0×105,8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1.0×104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1.0×103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.

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

0bjective 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 1×103,1×104,0.5×105,1×105 and 5×105 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 Tis-sues of liver,spleen,lungs,or cells from peripheral blood,fight tibia,right femur,left tibia and left femur24 hours after xenotransplantation in iBM injecting 5×105 CD34+ cells recipients expressed human chromo-some 17 specific α-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 a11 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 1×105 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 1.0×104CD34+ cells(P<0.01).Human cell engraftment was observed in iBM but not in IV recipients when transplanted with 1.0×103 CD34+ cells.and was usually obsened in non-injected bones.Conclusion Intra-bone marrow strategy can efficiently in-crease the engraftment of umbilical cord blood derived hematopoietic stem/progenitor cells in xenotransplanted NOD/SCID mouse model.