Human monocytes differentiate into macrophages under the influence of human KPB-M15 conditioned medium

Culture medium conditioned by the L929 murine fibroblast cell line contains macrophage colony-stimulating factor (M-CSF), providing an alternative to recombinant M-CSF for in vitro generation of murine macrophages. No such alternative has been described for in vitro studies requiring human macrophages. We tested the differentiation of human blood monocytes into mature macrophages by culturing in media conditioned by the human KPB-M15 cell line, which produces M-CSF and interleukin 6 (IL-6). The phenotypes of macrophages cultured in KPB-M15 conditioned media and recombinant M-CSF were compared by examining viability, expression of cell surface markers, phagocytic/pinocytic activity, and cytokine/chemokine secretion in response to bacterial lipopolysaccharide (LPS). In conditioned medium, monocytes differentiated into a homogeneous population of large cells that exhibited higher expression of CD14 and the macrophage mannose receptor (CD206) than did M-CSF-cultured cells. Cells matured in KPB-M15 conditioned medium exhibited macrophage morphology, were phagocytic, and were activated in response to LPS. These data demonstrate that KPB-M15 conditioned medium can be used to differentiate human blood monocytes into macrophages in vitro.