TY - JOUR
T1 - Synergism between erythropoietin and interleukin-3 in the induction of hematopoietic stem cell proliferation and erythroid burst colony formation
AU - Migliaccio, G.
AU - Migliaccio, A. R.
AU - Visser, J. W.M.
PY - 1988
Y1 - 1988
N2 - 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-fluourouracil-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 (≤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.
AB - 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-fluourouracil-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 (≤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.
UR - http://www.scopus.com/inward/record.url?scp=0023758312&partnerID=8YFLogxK
U2 - 10.1182/blood.v72.3.944.944
DO - 10.1182/blood.v72.3.944.944
M3 - Article
C2 - 3262001
AN - SCOPUS:0023758312
SN - 0006-4971
VL - 72
SP - 944
EP - 951
JO - Blood
JF - Blood
IS - 3
ER -