JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
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Presence of osteoclast precursors in colonies cloned in the presence of hematopoietic colony-stimulating factors.

Osteoclasts are derived from hematopoietic stem cells, but the relationship between osteoclast precursors (OCPs) and hematopoietic colony-forming cells (CFCs) has not yet been clarified. Although osteoclasts share certain cell surface markers and growth factor requirements with their macrophage and monocyte cell lineages, osteoclasts are a different lineage with regard to the requirement for signaling via c-Kit. To investigate whether CFCs are able to differentiate into osteoclasts, we performed in vitro studies of osteoclastogenesis. We performed progenitor assays in the presence of hematopoietic colony-stimulating factors. Primary colonies were plucked and examined for their potential to differentiate into osteoclasts. We found that osteoclasts are present in colonies elicited by macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor kB ligand (RANKL) in semisolid cultures. Moreover, a part of the cells composing the colonies elicited by granulocyte-macrophage colony-stimulating factor (GM-CSF) or M-CSF alone possessed the potential to differentiate into osteoclasts. These OCPs in the colonies were enriched in the c-Fms+ large-sized cell fraction and had a foamy cell morphology, like mature macrophages. A small number of cells in M-CSF-promoted and GM-CSF-promoted colonies formed secondary colonies in the semisolid medium containing these factors. The frequency of OCPs in these secondary colonies elicited by M-CSF was 10 times higher than that elicited by GM-CSF. Multiple origins of OCPs that differentiate into mature osteoclasts are proposed based on the observation that osteoclasts could be generated from OCPs that emerged from CFCs induced under different conditions or developmental stages.

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