阵发性睡眠性血红蛋白尿症CD34^+CD59^+细胞长期造血的动物实验  

Long-term ability for hematopoiesis of CD34^+ CD59^+ cells from patients with paroxysmal nocturnal hemoglobinuria:experiment with mice

作　　者：孙婉玲[1,2] 韩冰[1] 汪玄[1] 钟玉萍[1] 庄俊玲[1] 沈悌[1] 武永吉[1] 

机构地区：[1]中国医学科学院 中国协和医科大学 北京协和医院血液科,100730 [2]首都医科大学宣武医院血液科工作

出　　处：《中华医学杂志》2007年第42期3012-3015,共4页National Medical Journal of China

摘　　要：目的获得阵发性睡眠性血红蛋白尿症(PNH)患者正常克隆干细胞(CD34^+CD59^+)支持长期造血的证据。方法纯化、扩增 PNH 患者骨髓 CD34^+CD59^+细胞并移植入联合免疫缺陷(SCID)小鼠体内,90 d 后以首次移植的小鼠骨髓细胞进行二次移植。结果分别移植了 PNH 患者和正常对照骨髓 CD34^+CD59^+细胞的小鼠在移植后第90天,外周血细胞水平均恢复正常;两组小鼠的组织中均有 CD45的表达,且差异无统计学意义。分别移植了由 PNH 患者和正常对照骨髓 CD34^+CD59^+细胞重建造血的小鼠骨髓细胞的小鼠在移植后第30天,外周血细胞水平差异无统计学意义;两组小鼠的组织中均有 CD45的表达,且差异无统计学意义;移植后小鼠骨髓细胞里均可扩增出供者的 SRY 基因片段。结论经体外纯化、扩增后的 PNH 患者骨髓 CD34^+CD59^+细胞具有与正常CD34^+CD59^+细胞同样的长期造血的能力。Objective To obtain the evidence of long-term hematopoiesis by normal clone stem cells from patients with paroxysmal nocturnal hemoglobinuria （PNH）. Methods 10 ml fresh bone marrow was collected from 2 PNH patients. Normal bone marrow was obtained from the ribs resected during operation of 2 patients without hematopathy. CD34^＋CD59^＋ cells were isolated, purified, and expended. 12 total body irradiated SCID mice were divided into 2 equal group to undergo transplantation of the expanded CD34^＋CD59^＋ cells from PNH patients （ experiment group 1 ） or normal patients （ control group 1 ）. Four mice underwent transfusion of cell-free medium as blank control group 1. 30 and 60 days after the transplantation peripheral blood samples were collected and 90 days after the transplantation heart blood samples were collected. Spleens were taken out to prepare suspension of splenocytes and bacterium-free bone marrow cell suspension was prepared from the bilateral femurs. 90 days after the primary transplantation another 16 mice underwent total body irradiation and then divided into 2 equal groups to undergo transplantation of the bone marrow cells of the mice of the experiment group 1 and control group 1 （ experiment group 2 and control group 2）. Another 4 mice were transfused with cell-free medium fluid （ blank control group 2）. The levels of blood cells of all groups were calculated 30, 60, and 90 days after the primary and 30 days after the secondary transplantation. The cell percentages of peripheral blood, spleen, and bone marrow after the primary transplantation and 30 days after the secondary transplantation were detected by flow cytometry. PCR was used to detect whether the sex determining region Y existed in thee female SCID mice after transplantation. Results Ninety days after primary transplantation, the peripheral blood cell levels of both experiment group 1 and control group 1 recovered to normal. Expression of CD45 was detected in the mice without significant difference between the 2 gr

关 键 词：血红蛋白尿 阵发性 CD34^+CD59^+’细胞 造血干细胞 小鼠 SCID 

分 类 号：R55[医药卫生—血液循环系统疾病]