小鼠胎肝单个核细胞行骨髓腔内注射重建造血系统的效果

Intra-bone marrow injection of fetal liver mononuclear cells to reconstruct the hematopoietic system in mice

目的·观察用小鼠胎肝单个核细胞悬液行骨髓腔内注射以重建小鼠造血系统的效果。方法·用Ficoll分离技术获取胎龄13.5 d的小鼠胎肝单个核细胞悬液,采用磁珠分选的方法获得造血干细胞(hematopoietic stem cell,HSC)并计数。以骨髓腔内注射的方法对137Cs辐射清髓后的小鼠胫骨进行小鼠胎肝单个核细胞移植。对移植后的受体小鼠通过血常规检测、血涂片及骨髓涂片镜检的方法观察造血系统的重建情况。对受体小鼠移植后1年骨髓中的供体HSC进行流式细胞检测。结果·胎龄13.5 d的小鼠胎肝单个核细胞中HSC占0.171%。小鼠胎肝单个核细胞移植后1周,外周血即能探测到供体细胞来源的血细胞;移植后第3周受体小鼠外周血白细胞、红细胞、血红蛋白基本恢复至辐照前水平;第5周外周血中供体细胞来源的血细胞比例趋于稳定;移植后1年受体小鼠骨髓中的HSC基本来自供体细胞。结论·小鼠胎肝单个核细胞通过骨髓腔内注射的移植方法能高效重建小鼠造血系统。

Objective To observe the effect of intra-bone marrow injection(IBMI) of fetal liver mononuclear cell suspension on reconstructing the hematopoietic system in mice. Methods The fetal liver mononuclear cells from the mice with gestational age of 13.5 days were obtained by Ficoll separation technique, and then the hematopoietic stem cells(HSCs) were obtained by magnetic bead sorting. The fetal liver mononuclear cells were injected into the tibias of 137 Cs irradiated mice by IBMI. The recipient mice were observed for reconstitution of the hematopoietic system by routine blood examination, blood smear and bone marrow smear. The donor HSCs in the bone marrow of the recipient mice was subjected to flow cytometry 1 year after transplantation. Results HSCs accounted for 0.171% in the mouse fetal liver mononuclear cells. One week after transplantation of mouse fetal liver mononuclear cells, blood cells derived from donor cells were detected from peripheral blood;in the third week after transplantation, peripheral blood leukocytes, red blood cells and hemoglobin of recipient mice were restored to the pre-irradiation levels;the percentage of donor-derived blood cells in peripheral blood in the fifth week was stable;the HSCs in the bone marrow of recipient mice were derived from donor cells 1 year after transplantation. Conclusion The murine fetal liver mononuclear cells can efficiently reconstruct the hematopoietic system of mice by IBMI.