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Type: Journal article
Title: The generation of osteoclasts from RAW 264.7 precursors in defined, serum-free conditions
Author: Vincent, C.
Kogawa, M.
Findlay, D.
Atkins, G.
Citation: Journal of Bone and Mineral Metabolism, 2009; 27(1):114-119
Publisher: Springer-Verlag Tokyo
Issue Date: 2009
ISSN: 0914-8779
Statement of
Cristina Vincent, Masakazu Kogawa, David M. Findlay and Gerald J. Atkins
Abstract: Osteoclasts are the unique cell type capable of resorbing bone. The discovery of the TNF-ligand family member, RANKL, has allowed more reliable study of these important cells. The mouse monocytic cell line, RAW 264.7, has been shown to readily differentiate into osteoclasts upon exposure to recombinant RANKL. Unlike primary osteoclast precursors, there is no requirement for the addition of macrophage colony stimulating factor (M-CSF). However, to date, their differentiation has always been studied in the context of added foetal calf serum (FCS). FCS is a complex and largely undefined mixture of growth factors and matrix proteins, and varies between batches. For this reason, osteoclastogenesis would ideally be studied in the context of a defined, serum-free medium. RAW 264.7 cells were cultured in serum-replete α-MEM or serum-deprived medium (SDM) shown previously to support the growth of human osteoclasts in a co-culture with normal osteoblasts. In SDM, in the presence of recombinant RANKL, RAW 264.7 cells readily differentiated into tartrate resistant acid phosphatase (TRAP) positive multinucleated osteoclast-like cells, a process that was enhanced with the addition of 1α,25-dihydroxyvitamin D3 (1,25D). While the osteoclasts grown in SDM were smaller in size compared with those derived in serum-replete media, their resorptive capacity was significantly increased as indicated by a twofold increase in average resorption pit size. In conclusion, we describe a defined model for studying osteoclast differentiation and activity in the absence of serum, which will be ideal for studying the role of agonistic and antagonistic molecules in this process.
Keywords: Osteoclast; Osteoclastogenesis; RAW 264.7; Differentiation; RANKL
RMID: 0020090025
DOI: 10.1007/s00774-008-0018-6
Appears in Collections:Orthopaedics and Trauma publications

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