Generation and functional characterisation of canine bone marrow-derived macrophages

Abstract

A culture of bone marrow cells from the femurs of canine pups at high concentrations of fetal calf serum under non-adherent conditions allowed the proliferation and differentiation of mononuclear phagocyte lineage cells, as evidenced by morphology and CD14 expression. Cells from other lineages progressively diminished in numbers. Cells collected between 12 and 19 days of culture expressed an array of macrophage activities including ingestion of opsonised erythrocytes, generation of superoxide, up-regulation of procoagulant activity and synthesis of tumour necrosis factor (TNF) upon appropriate stimulation. TNF production was enhanced when the cultures were simultaneously stimulated with canine recombinant, or supernatant-derived, interferon-gamma. In contrast, low levels of inducible nitric oxide (NO) synthase were expressed by only a minority of stimulated macrophages, and nitrite could not be detected in the medium. Therefore, canine macrophages generated by this novel culture system resemble human macrophages in their inefficient and restricted generation of NO upon appropriate stimulation.