含脐血MSCs体系扩增后的脐血重建NOD／SCID小鼠造血的研究  

NOD/SCID Mice Hematopoietic Repopulating Ability of ex vivo Expanded Umbilical Cord Hematopoietic Blood Cells which Co-cultured with MSCs from Human Umbilical Cord Blood

作　　者：周敦华[1] 张绪超[1] 黄绍良[1] 方建培[1] 吴燕峰[1] 魏菁[1] 包蓉[1] 黄科[1] 黎阳[1] 

机构地区：[1]中山大学附属第二医院儿科,广东广州510120

出　　处：《中山大学学报（医学科学版）》2005年第6期639-643,691,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金(00137)广东省卫生厅基金(E200040)

摘　　要：【目的】探讨含人脐血(umbilical cord blood，UCB)来源的间充质干细胞(mesenchymal stem cells，MSCs)联合造血生长因子(hematopoietic growth factors，HGFs)体系扩增后的脐血造血细胞在NOD／SCID小鼠体内植入及重建小鼠造血的能力。【方法】①用含女性胎儿脐血MSCs(UCBMSCs)及HGFs组合的无血清扩增体系体外扩增男性新生儿UCB CD34^+细胞。②收获扩增第6天的UCB细胞，经尾静脉移植给亚致死量照射后的雌性NOD／SCID小鼠。③动态观察移植后小鼠生存情况及外周血象的恢复。④移植后8周，流式细胞仪检测存活小鼠骨髓中人CD45^+、CD45^+CD33^+、CD45^+CD41^+、CD45^+CD3^+和CD45^+CD19^+细胞含量；PCR法检测移植小鼠外周血中人Y染色体表达。【结果】①非扩增组小鼠存活率为60％，扩增组存活率为90％。②动态观察发现：扩增组白细胞和血红蛋白于移植后第20天恢复，PLT于移植后第40天恢复，明显短于非扩增组。③移植后8周，两组小鼠外周血中人Y染色体检测均为阳性，扩增组和非扩增组小鼠骨髓中人CD45+细胞的含量分别为18.5％±8.3％和16.5％±5.7％，差异无统计学意义(P＞0.05)。④移植后8周，扩增组小鼠体内CD45^+CD33^+和CD45^+CD41a^+细胞的百分率均高于非扩增组，但CD45^+CD3^+、CD45^+CD19^+细胞百分比均低于非扩增组。【结论】①含人UCBMSCs体系扩增后的UCB细胞具有NOD／SCID小鼠体内植入并重建小鼠造血的能力，似不具有提高小鼠体内植入率的作用。②扩增后的UCB移植可显著促进移植后小鼠造血恢复，提高小鼠存活率。③扩增后的UCB移植主要促进小鼠髓系及巨核系的植入，但对淋巴系的植入有抑制作用。[Objective] To evaluate the engraftment and hematopoietic reconstruction ability of ex vivo expanded umbilical cord blood （UCB） hematopoietic cells which co-cultured with MSCs from human umbilical cord blood combind with hematopoietic growth factors （HGFs） in NOD/SCID mice. [Methods] （1） Human male UCB CD34^＋ cells were incubated in the system with contains female UCBMSCs, HGFs and serum free medium. （2） The expanded UCB hematopoietic cells on the 6th day were infused into sublethally irradiated （250 cGy） NOD/SCID mice through lateral tail vein injection. （3） Existence state and hematopoietic recover of transplanted mice were detected dinamically. （4） On the 8th week after transplantation, human derived CD45^＋, CD45^＋CD34^＋, CD45^＋CD3^＋, CD45^＋CD19^＋, CD45^＋CD33^＋ cells in the BM of mice were detected by flow cytometry assay and human Y chromosome was detected in the peripheral blood by PCR method. [Results] （1） The survival rate of un-expanded group and expanded group were 60% vs 90%. （2） The expanded group had rapid recovery in WBC, hemoglobin, and platelet. （3） Eight weeks after transplantation, human Y chromosomes were all positive in transplanted mice, and there was no significant difference （P 〉0.05） between control and test groups in the percent of human CD45^＋ cells in mice BM. （4） Eight weeks after transplantation, the CD45^＋CD33^＋ cells and CD45^＋CD41a^＋ cells were significantly higher in expanded group than that in unexpanded group, and CD45^＋CD3^＋ cells and CD45^＋CD19^＋ cells were significantly lower in expanded group. [Conclusion]（1） Expanded UCB hematopoietic cells can reconstitute the hematopoiesis in radiated NOD-SCID mice, but it can not increase engraftment rate. （2） Expanded UCB hematopoietic cells can inhance the recovery of hematopoiesis reconstitution and improved the longevity of transplanted mice. （3） Expanded UCB transplantation mainly enhanced the engraftment of granulocytes and megacaryoc

关 键 词：间充质干细胞 植入 造血重建 NOD/SCID小鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]