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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 evidence of long-term hematopoiesis in normal clonal stem cells (CD34 ~ + CD59 ~ +) in patients with paroxysmal nocturnal hemoglobinuria (PNH). Methods The bone marrow CD34 ~ + CD59 ~ + cells were purified and expanded in PNH patients and were transplanted into SCID mice. After 90 days, the bone marrow cells were transplanted for the first time in mice. Results The levels of CD45 + CD59 + cells in peripheral blood of PNH patients and normal control mice were all restored to normal levels on the 90th day after transplantation. The expression of CD45 in both groups was significantly different Statistical significance. There was no significant difference in the level of peripheral blood cells in mice transplanted with bone marrow cells reconstituted from PNH patients and normal control bone marrow CD34 ~ + CD59 ~ + cells on day 30 after transplantation. In the tissues of both groups All had CD45 expression, and the difference was not statistically significant. The donor SRY gene fragment could be amplified in the mouse bone marrow cells after transplantation. Conclusion The purified CD34 + CD59 + cells from PNH patients after in vitro purification have the same long-term hematopoiesis ability as normal CD34 + CD59 + cells.