In cultures of spleen cells from normal mice, recombinant human granulocyte colony-stimulating factor (G-CSF) supported the formation of multipotential blast cell colonies. Serial replating of the blast cell colonies in the presence of G-CSF, however, failed to demonstrate any direct effect of G-CSF on murine multipotential progenitors. We therefore examined the effects of G-CSF in combination with murine interleukin 3 on proliferation of murine blast cell colony-forming cells. The time course of total colony formation and multilineage colony formation by spleen cells harvested from mice 4 days after injection of 5-fluorouracil at 150 mg/kg was significantly shortened in cultures containing both factors in contrast with cultures supported by either factor alone. Serial observations of individual multipotential blast cell colonies (mapping) revealed that blast cell colonies emerged at random time intervals in the presence of interleukin 3 or G-CSF. The appearance of blast cell colonies, however, was significantly hastened in cultures containing both factors relative to cultures grown with either factor. In cultures of day-2 post-5-fluorouracil bone marrow cells, G-CSF in concentrations as low as 1 unit/ml revealed synergism with interleukin 3 in supporting the proliferation of multipotential progenitors. This synergistic activity may explain the previous in vivo studies suggesting the effects of G-CSF on apparent multipotential stem cells
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