Abstract
Bone homeostasis depends on balanced bone deposition and bone resorption, which are mediated by osteoblasts and osteoclasts, respectively. The process of bone turnover requires the coordination of these cells. Changes in the ability of either cell type to perform its function results in pathological conditions such as osteoporosis and tumor-induced bone loss (osteolysis). The number of osteoclasts present at the site of bone remodeling as well as the activity of those osteoclasts the control amount of bone resorbed (1). Therefore, factors affecting overall numbers of osteoclasts and osteoclast activation are key to regulating bone loss. Osteoclast numbers are in part controlled by osteoclast differentiation from bone marrow precursors of the monocyte/macrophage lineage (2). Differentiation of these hematopoietic precursors into osteoclasts is supported by bone marrow stromal cell production of two cytokines, receptor activator of NF-κB ligand (RANKL) and macrophage colony stimulating factor (M-CSF), which are both necessary and sufficient to mediate osteoclast differentiation (3, 4). Although RANKL production by the stroma supports osteoclast differentiation, this process is antagonized by osteoprotogerin (OPG) production, which acts as a soluble decoy receptor for RANKL (5, 6). Mechanistic studies to elucidate the factors influencing bone metabolism necessitate in vitro studies of osteoclast differentiation, activation and survival. There are a number of in vitro methods used to culture and study osteoclasts, some of which are described in this chapter.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
1. Suda, T., et al. (1997) Regulation of osteoclast function. J Bone Miner Res 12, 869–879.
2. Udagawa, N., et al. (1990) Origin of osteoclasts: mature monocytes and macrophages are capable of differentiating into osteoclasts under a suitable microenvironment prepared by bone marrow-derived stromal cells. Proc Natl Acad Sci U S A 87, 7260–7264.
3. Arai, F., et al. (1999) Commitment and differentiation of osteoclast precursor cells by the sequential expression of c-Fms and receptor activator of nuclear factor kappaB (RANK) receptors. J Exp Med 190, 1741–1754.
4. Wong, B. R., et al. (1997) TRANCE is a novel ligand of the tumor necrosis factor receptor family that activates c-Jun N-terminal kinase in T cells. J Biol Chem 272, 25190–25194.
5. Simonet, W. S., et al. (1997) Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell 89, 309–319.
6. Yasuda, H., et al. (1998) Osteoclast differentiation factor is a ligand for osteoprotegerin/osteo-clastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A 95, 3597–3602.
7. Lacey, D. L., et al. (1998) Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93, 165–176.
8. Udagawa, N., et al. (1989) The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclast-like cell differentiation in cocultures with mouse spleen cells. Endocrinology 125, 1805–1813.
9. Takahashi, N., et al. (1988) Osteoblastic cells are involved in osteoclast formation. Endocrinology 123, 2600–2602.
10. Raschke, W. C., et al. (1978) Functional macrophage cell lines transformed by Abelson leukemia virus. Cell 15, 261–267.
11. Sells Galvin, R. J., et al. (1999) TGF-beta enhances osteoclast differentiation in hematopoietic cell cultures stimulated with RANKL and M-CSF. Biochem Biophys Res Commun 265, 233–239.
12. Ragab, A. A., et al. (1998) Osteoclast differentiation requires ascorbic acid. J Bone Miner Res 13, 970–977.
13. Chambers, T. J., et al. (1984) Resorption of bone by isolated rabbit osteoclasts. J Cell Sci 66, 383–399.
14. Boyde, A., et al. (1986) Scanning electron microscopy in bone pathology: review of methods, potential and applications. Scan Electron Microsc Pt 4, 1537–1554.
15. Onyia, J. E., et al. (2004) Novel and selective small molecule stimulators of osteoprotegerin expression inhibit bone resorption. J Pharmacol Exp Ther 309, 369–379.
16. Fukayama, S., et al. (1988) Human parathyroid hormone (PTH)-related protein and human PTH: comparative biological activities on human bone cells and bone resorption. Endocrinology 123, 2841–2848.
Acknowledgments
The authors thank R. Galvin and E. Greenfield for their helpful insight. Funding sources are NIH grants
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bradley, E.W., Oursler, M.J. (2008). Osteoclast Culture and Resorption Assays. In: Westendorf, J.J. (eds) Osteoporosis. Methods In Molecular Biology™, vol 455. Humana Press. https://doi.org/10.1007/978-1-59745-104-8_2
Download citation
DOI: https://doi.org/10.1007/978-1-59745-104-8_2
Publisher Name: Humana Press
Print ISBN: 978-1-58829-828-7
Online ISBN: 978-1-59745-104-8
eBook Packages: Springer Protocols