分化抑制培养体系支持小鼠骨髓造血细胞体外扩增、归巢及造血重建

Support of ex vivo expansion, homing and hematopoiesis reconstitution by differentiation inhibiting culture system in hematopoietic cells from mouse marrow

目的探讨分化抑制培养体系对小鼠骨髓造血细胞体外扩增、归巢及造血重建的支持作用。方法将小鼠骨髓造血细胞在分化抑制培养体系中共培养7 d,检测细胞总数、集落形成细胞(CFC)及干细胞抗原1阳性(Sca-1+)细胞,了解细胞的扩增情况。将扩增后的雄性小鼠(供者)骨髓造血细胞移植给雌性小鼠(受者),移植后24 h,用FISH方法检测受者体内的供者Y染色体阳性细胞归巢率,并在移植后第3、6、9、12、15、18、25、32、42 d,分别检测受者的红细胞、白细胞及血小板,了解造血功能变化。结果分化抑制培养体系能够在体外扩增小鼠骨髓造血细胞,细胞总数、CFC及Sca-1+细胞明显增加,扩增后的细胞其归巢率增加,能使受者外周血的红细胞、白细胞及血小板恢复加快。结论分化抑制培养体系支持小鼠骨髓造血细胞体外扩增、归巢及造血重建,显示它是一种良好的扩增体系。

Objective To explore whether differentiation inhibiting culture system (DICS) supports ex vivo expansion, homing and hematopoiesis reconstitution of hematopoietic cells from mouse marrow. Methods DISC were co- cultured with hemapoietic cells from mouse marrow for 7 days. The total number of cells, CFC and Sca-1 + cells were counted. The expanded cells from male donor were transplanted. To female recipients, the homing percentage of Y-chromosome from male donor in FISH was measured 24 h after transplantation. The number of RBC, WBC and PLT in peripheral blood was counted 3, 6, 9, 12, 15, 18, 25, 32 and 42 days after transplantatioa Results The total number of cells, CFC and Sca-1+ cells were significantly increased. The homing of expanded cells was increased, and the number of RBC, WBC and PLT was recovered more quickly. Conclusion Differentiation inhibiting culture system (DICS) can support ex vivo expansion, homing and hematopoiesis reconstitution of hematopoietic cells from mouse marrow, and it is a good expansion system.