When spleen cells of 5-fluorouracil (5-FU)-treated mice were cultured in the presence of interleukin 3 (IL-3), most colonies revealed IL-3 concentration-dependent colony formation except for mast cell colonies and blast cell colonies. While most colonies were smaller in lower concentrations of IL-3, the size of the blast cell colonies were similar between high and low IL-3 groups. These data suggested that blast cell colony development requires less IL-3 than the development of multilineage colonies from blast cell colonies. This notion was supported by experiments in which IL-3 was added twice to cultures of spleen cells of 5-FU-treated mice. When low concentrations of IL-3 were added on day 7, there was a reduction in the number of multilineage colonies formed without an effect on the number of blast cell colonies. Using this information, we developed a culture system that favors blast cell colony formation by cells of normal mice. When low (20 U/ml) concentrations of IL-3 were added to cultures of spleen cells of normal mice on day 7 of incubation in media containing 2-5% fetal calf serum, blast cell colonies were the predominant colony type. The blast cell colonies revealed high but variable secondary replating efficiencies. These data suggest that multipotential progenitors may become less sensitive to IL-3 as they differentiate in culture.
K Koike, J N Ihle, M Ogawa