Abstract
Osteoclasts derive from macrophage colony-stimulating factor (M-CSF)-dependent hemopoietic precursors that develop into cells that express the αvβ3 subunit of the vitronectin receptor (VNR) and the calcitonin receptor (CTR). The extracellular degradative process, known as bone resorption, is the hallmark of the osteoclast, and includes removal of both the hydroxyapatite and organic components of the skeleton. For bone resorption to occur, osteoclasts form a subcellular space, referred to as an extracellular lysosome, into which they secrete acid and enzymes when they come into contact with either calcified bone or dentine but not with plastic or uncalcified collagen-based matrices. This subcellular space is dependent upon the formation of a “tight seal” by the osteoclast, a process involving rearrangement of the cytoskeleton into a characteristic F-actin ring structure (see the chapter by Nesbitt and Horton, this volume, for details).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Flanagan, A. M., Sarma, U., Steward, C. G., Vellodi, A., and Horton M. A. (2000) Study of the non-resorptive phenotype of osteoclast-like cells from patients with malignant osteopetrosis: a new approach to investigating pathogenesis. J. Bone Miner. Res. 15, 1–9.
Lader, C. S., Scopes, J., Horton, M. A., and Flanagan, A. M. (2001) Generation of human osteoclasts in stromal cell-free and stromal cell-rich cultures: differences in osteoclast CD11c/CD18 integrin expression. Br. J. Haematol. 111, 1210–1217.
Sarma, U., Edwards, M., Motoyoshi, K., and Flanagan, A. M. (1998) 17β-Estradiol inhibits human osteoclast formation in vitro. J. Cell. Physiol. 175, 99–108.
Lader, C. S. and Flanagan, A. M. (1998) Prostaglandin E2, interleukin 1α and tumor necrosis factor α increase human osteoclast formation and bone resorption in vitro. Endocrinology 139, 3157–3164.
Massey, H. M., Scopes, J., Horton, M. A., and Flanagan, A. M. (2001) Transforming growth factor-β 1 stimulates the osteoclast-forming potential of the haemopoietic precursor in peripheral blood cells in a lymphocyte-rich microenvironment. Bone 28, 577–582.
Scopes, J., Massey, H. M., Ebrahim, H., Horton, M. A., and Flanagan, A. M. (2001) Interleukin-4: bidirectional effects on human osteoclast formation. Bone 29, 203–208.
Fujikawa, Y., Quinn, J. M. W., Sabokbar, A., McGee, J. O. D., and Athanasou, N. A. (1996) The human osteoclast precursor circulates in the monocyte fraction. Endocrinology. 137, 4058–4060.
Massey, H. M. and Flanagan A. M. (1999) Human osteoclasts derive from CD14-positive monocytes. Br. J. Haematol. 106, 167–170.
Kong, Y.-Y., Yoshida, H., Sarosi, I., et al. (1999) OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature 397, 315–323.
Kong, Y. Y., Feige, U., Sarosi, L., et al. (1999) Activated T cells regulate bone loss and joint destruction in adjuvant arthritis through osteoprotegerin ligand. Nature 402, 304–308.
Sells Galvin, R. J., Gatlin, C. L., Horn, J. W., and Fuson, T. R. (1999) TGF-β enhances osteoclast differentiation in hematopoietic cell cultures stimulated with RANKL and M-CSF. Biochem. Biophys. Res. Commun. 265, 233–239.
Fuller, K., Lean, J. M., Bayley, K. E., Wani, M. R., and Chambers, T. J. (2000) A role for TGFβ(1) in osteoclast differentiation and survival. J. Cell Sci. 113, 2445–2453.
Sarma, U. and Flanagan, A. M. (1996) Macrophage-colony stimulating factor (M-CSF) induces substantial osteoclast formation in human bone marrow cultures. Blood 88, 2531–2540.
Suda, T., Takahashi, N., Udagawa, N., Jimi, E., Gillespie, M. T., and Martin, T. J. (1999) Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr. Rev. 20, 345–357.
Lea, C. K., Sarma, U., and Flanagan, A. M. (1999) Macrophage colony stimulating-factor transcripts are differentially regulated in rat bone-marrow by gender hormones. Endocrinology 140, 273–279.
Flanagan, A. M. and Lader, C. S. (1998) Update on the biological effects of macrophage colony-stimulating factor. Curr. Opin. Hematol. 5, 181–185.
Fuller, K., Wong, B., Fox, S., Choi, Y., and Chambers, T. C. (1998) TRANCE is necessary and sufficient for osteoblast-mediated activation of bone resorption in osteoclasts. J. Exp. Med. 188, 997–1001.
Tobias, J. and Chambers, T. J. (1989) Glucocorticoids impair bone resorptive activity and viability of osteoclasts disaggregated from neonatal rat long bones. Endocrinology 125, 1290–1295.
Matsukaki, K., Udagawa, N., Takahashi, N., et al. (1998) Osteoclast differentiation factor (ODF) induces osteoclast-like cell formation in human peripheral blood mononuclear cell cultures. Biochem. Biophys. Res. Commun. 246, 199–204.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Flanagan, A.M., Massey, H.M. (2003). Generating Human Osteoclasts In Vitro from Bone Marrow and Peripheral Blood. In: Helfrich, M.H., Ralston, S.H. (eds) Bone Research Protocols. Methods in Molecular Medicine, vol 80. Humana Press. https://doi.org/10.1385/1-59259-366-6:113
Download citation
DOI: https://doi.org/10.1385/1-59259-366-6:113
Publisher Name: Humana Press
Print ISBN: 978-1-58829-044-1
Online ISBN: 978-1-59259-366-8
eBook Packages: Springer Protocols