Migliaccio G, Migliaccio A R, Visser J W
Department of Hematology, Istituto Superiore Sanita, Rome.
Blood. 1988 Sep;72(3):944-51.
The influence of recombinant erythropoietin (Ep) and interleukin-3 (IL-3) on the proliferation and differentiation of murine hematopoietic stem and progenitor cells was investigated in serum-deprived cultures. The differentiation of progenitor cells, purified by collecting blast cell colonies from spleen cell cultures of 5-fluorouracil-treated mice, was evaluated by scoring the number and type of colonies appearing after eight days in semisolid culture. IL-3 induced the formation of both erythroid and granulocyte-macrophage colonies in a concentration-dependent fashion, the plateau being reached at 300 U/mL. However, concentrations of IL-3 alone that had little or no effect (less than or equal to 10 U/mL) induced maximal numbers of erythroid bursts in the presence of Ep (1.5 IU/mL). In the presence of Ep alone, no colonies were seen. Proliferation of quiescent hematopoietic stem cells, purified by cell sorting and evaluated by spleen colony assay (CFU-S), was investigated by measuring the total cell number and CFU-S content and the DNA histogram at 20 and 48 hours of liquid culture. Almost no cells or CFU-S survived 20 hours of incubation without the addition of IL-3. The presence of either IL-3 (400 U/mL) or the combination of EP and IL-3 (10 U/mL), supported the maintenance of nearly 40% of sorted CFU-S for 48 hours. Approximately 10% of these cells were in the S phase of the cell cycle at 20 hours and an increase in the total cell number per culture, but not in the CFU-S content, was detected at 48 hours. These data indicate that IL-3 exerts a differentiative and proliferative effect on early stem and progenitor cells, which is concentration dependent. At IL-3 concentrations, which had little or no activity alone, Ep acted synergistically to induce both proliferation of stem cells and differentiation of erythroid progenitors.
在血清饥饿培养中研究了重组促红细胞生成素（Ep）和白细胞介素-3（IL-3）对小鼠造血干细胞和祖细胞增殖及分化的影响。通过从5-氟尿嘧啶处理小鼠的脾细胞培养物中收集原始细胞集落来纯化祖细胞，通过对半固体培养八天后出现的集落数量和类型进行评分来评估其分化情况。IL-3以浓度依赖方式诱导红系和粒-巨噬细胞集落的形成，在300 U/mL时达到平台期。然而，单独使用IL-3时几乎没有或没有作用的浓度（小于或等于10 U/mL），在存在Ep（1.5 IU/mL）时诱导出最大数量的红系爆式集落。单独存在Ep时，未观察到集落。通过细胞分选纯化并通过脾集落测定（CFU-S）评估的静止造血干细胞的增殖，通过在液体培养20小时和48小时时测量总细胞数、CFU-S含量和DNA直方图来进行研究。在不添加IL-3的情况下，孵育20小时后几乎没有细胞或CFU-S存活。IL-3（400 U/mL）或Ep与IL-3（10 U/mL）的组合的存在，支持近40%的分选CFU-S维持48小时。这些细胞中约10%在20小时时处于细胞周期的S期，并且在48小时时检测到每个培养物中总细胞数增加，但CFU-S含量未增加。这些数据表明，IL-3对早期干细胞和祖细胞发挥分化和增殖作用，且具有浓度依赖性。在单独作用几乎没有或没有活性的IL-3浓度下，Ep发挥协同作用，诱导干细胞增殖和红系祖细胞分化。
