Human multilineage colony-stimulating factor (multi-CSF)/interleukin-3 (IL-3) induces colony formation from CFU-GEMM, BFU-E, and CFU-Eo when applied to in vitro cultures of highly enriched hematopoietic progenitor cells. No granulocytic colonies are formed in response to IL-3. However, with appropriate assays, we demonstrate that IL-3 increases the size of G-CSF-induced granulocytic colonies; these colonies contain greater proportions of immature cells as compared with colonies stimulated by G-CSF alone. Furthermore, IL-3 promotes the survival of CFU-G in vitro, whereas in cultures not supplemented with IL-3, CFU-G extinguish within seven days. We conclude that IL-3, although it does not stimulate granulocytic colony formation by itself, regulates the survival and proliferative rate of granulocytic progenitors.