Nishi N, Nakahata T, Koike K, Takagi M, Naganuma K, Akabane T
Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Japan.
Blood. 1990 Oct 1;76(7):1330-5.
The effects of recombinant human erythropoietin (rEp) on murine hematopoietic progenitors were studied using a serum-free culture. A high concentration of rEp stimulated the formation of mixed erythroid-megakaryocyte colonies (EM colonies) and blast cell colonies, as well as erythroid colonies, erythroid bursts, and megakaryocyte colonies from normal mouse bone marrow cells. Direct effects of rEp on EM colony, megakaryocyte colony, and erythroid burst formation were confirmed by depletion of accessory cells such as T cells, B cells, and macrophages from crude bone marrow cells, and inhibition of the colonies by the addition of rabbit anti-rEp antibody to the culture in a dose-dependent fashion. Replating experiments were performed to confirm the differentiating ability of blast cell colonies grown in the presence of rEp. Most of the blast cell colonies yielded not only secondary erythroid colonies but also megakaryocyte colonies in the presence of 2 IU/mL rEp. Some of the blast cell colonies produced secondary EM colonies in the presence of 16 IU/ml rEp of 2 IU/mL rEp plus interleukin-3, although no granulocyte-macrophage colonies were found in the secondary culture. These results suggest that Ep acts not only as a late-acting factor that is specific for erythroid progenitors, but also as a bipotential EM-stimulating factor for murine hematopoietic cells.
使用无血清培养研究重组人促红细胞生成素（rEp）对小鼠造血祖细胞的影响。高浓度的rEp刺激正常小鼠骨髓细胞形成混合红系 - 巨核细胞集落（EM集落）、原始细胞集落以及红系集落、红系爆式集落和巨核细胞集落。通过从粗制骨髓细胞中去除辅助细胞（如T细胞、B细胞和巨噬细胞），以及以剂量依赖方式向培养物中添加兔抗rEp抗体来抑制集落，证实了rEp对EM集落、巨核细胞集落和红系爆式集落形成的直接作用。进行传代实验以确认在rEp存在下生长的原始细胞集落的分化能力。在2 IU/mL rEp存在的情况下，大多数原始细胞集落不仅产生次级红系集落，还产生巨核细胞集落。在16 IU/ml rEp或2 IU/mL rEp加白细胞介素 - 3存在的情况下，一些原始细胞集落产生次级EM集落，尽管在次级培养中未发现粒细胞 - 巨噬细胞集落。这些结果表明，Ep不仅作为红系祖细胞特异的晚期作用因子，而且作为小鼠造血细胞的双潜能EM刺激因子。
