小鼠异基因骨髓移植归巢阶段供鼠造血细胞的体内追踪

Donor Hematopoietic Cell Tracking In Vivo at the Homing Phase of Allo-Bone Marrow Transplantation in Mice

经静脉途径输注的造血细胞能通过血液循环归巢于骨髓造血组织并重建造血 ,但关于输注的造血干 /祖细胞在造血器官内的寄居比例或种植效率 ,尚不完全清楚。为探讨输注的造血细胞的组织器官分布特性 ,本研究使用PKH 2 6荧光染料一种标记于细胞膜上的红色荧光染料 ,标记于近交系小鼠的造血细胞表面 ,成功地建立了一种稳定可靠的造血细胞体内追踪方法 ,用流式细胞术和荧光显微镜分别定量、定性检测标记小鼠的造血细胞。应用该方法对小鼠异基因骨髓移植模型归巢阶段的供鼠早期骨髓细胞Sca 1+ 亚群细胞的去向及其在造血器官 (骨髓、脾 )和非造血器官 (肺、肝 )的分布 ,分别进行了体内追踪观察 ,结果显示 :①静脉输注的Sca 1+ 亚群在肺内短暂滞留。②静脉输入小鼠骨髓细胞后 60小时内 ,在受鼠的造血器官和非造血器官均有Sca 1+ 亚群骨髓细胞的寄居 ,其中骨髓的造血细胞寄居数量显著高于非造血器官的造血细胞寄居数量 (P <0 .0 5 )。本研究结果证明 :在骨髓移植归巢阶段 ,受鼠的非造血器官 (肺 ,肝等 )中 。

It has been well-known that intravenously infused hematopoietic stem and progenitor cells can home to the bone marrow and reconstitute hematopoiesis. However, little is understood about the homing efficiency or percentage of infused stem and progenitor cells. In order to examine distribution pattern of infused hematopoietic cells in the organs and tissues, a direct assay system to trace transplanted cells in vivo by employing PKH-26, a red fluorescent membrane dye, to label hematopoietic cells in inbred strain of mice transplanted cells (stem cell antigen-1 positive subpopulation cell, Sca-1 + cells)was introduced. The numbers of labeled cells was measured by means of flow cytometry and fluorescence microscopy. The early fate of infused Sca-1 + donor bone marrow cells after intravenous administration in a allogeneic mouse model was examined. The presence of infused donor cells with the fluorescent dye PKH-26 was evaluated within 60 hours in hematopoietic organ(bone marrow and spleen) and non-hematopoietic organ(lungs and liver) of recipients. The data showed that ① Following intravenous infusion, Sca-1 + donor bone marrow cells were detained in lungs shortly. ② Sca-1 + donor bone marrow cells localized to both hematopoietic organ(bone marrow and spleen) and non-hematopoietic organ(lungs and liver) for periods of up to 60 hours following infusion, however, the number of donor hematopoietic cells localized to bone marrow was more than that localized to non-hematopoietic organ (P<0.05). These results indicated that there were also donor early hematopoietic cells in non-hematopoietic organ of recipients at the homing phase in allo-BMT mice.