In vitro induction of swine peripheral blood monocyte proliferation by the fibroblast-derived murine hematopoietic...

The addition of conditioned medium from murine L929 fibroblasts (MGF) to cultures of swine peripheral blood mononuclear cells (MNL) resulted in growth of cells of macrophage/monocyte lineage (MO). Glass-adherent swine MNL, shown to be greater than 95% phagocytic MO, grew in the presence of MGF, whereas swine blood granulocytes and lymphocytes were not MGF-responsive. Primary and secondary MO growth were directly dependent on MGF presence and concentration. MGF-stimulated MO synthesized DNA, as measured by cellular incorporation of tritium-labeled thymidine (3H-TdR). This mitogenic response was maximal by 5 to 6 days in primary MO cultures and declined thereafter to a lower magnitude in secondary MO cultures. In the presence of MGF, viable MO numbers increased with an approximate population doubling time of 5 to 7 days in primary culture. This growth rate was prolonged, to about 10 to 12 days, for MGF-stimulated MO in secondary cultures. MGF removal from primary and secondary MO cultures resulted in rapid growth cessation and cell death. MGF-stimulated MO could not be sustained in secondary culture beyond 7 weeks. MGF-cultured MO were positive for latex phagocytosis, non-specific esterase, Fc-receptor expression, and could mediate antibody-dependent cell-mediated cytotoxicity. The MO-mitogenic principle of MGF was identified as the murine, macrophage-specific colony-stimulating factor, CSF-1 (M-CSF). The swine MO-proliferative response to MGF was inhibited by addition of monospecific goat antisera to M-CSF. Purified M-CSF stimulated the growth of swine MO from cultures of MNL and primary glass-adherent MO.