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Isolated Osteoclast Cultures

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Bone Research Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 80))

Abstract

Osteoclasts are large, multinucleated cells formed by the fusion of hematopoietic, mononuclear progenitors of the monocyte/macrophage lineage, and are the cells responsible for resorbing bone. Osteoclasts are usually few in number relative to other cell types in bone and are difficult to isolate because they are contained in a hard tissue; in addition, they are at the end of their proliferation and differentiation cycle, presenting problems for the creation of osteoclast cell lines. However, with the development, almost 20 years ago, of in vitro resorption pit formation models, using isolated primary, mature osteoclasts and mineralized bone or dentine matrix as a substrate (1,2), considerable progress was made in our understanding of osteoclast biology. Data from such short term cultures complements that obtained from bone organ culture resorption models and long-term cultures of osteoclast forming hematopoietic stem cells derived from marrow or peripheral blood.

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References

  1. Boyde, A., Ali, N. N., and Jones, S. J. (1984) Resorption of dentine by isolated osteoclasts in vitro. Br. Dent. J. 156, 216–220.

    Article  PubMed  CAS  Google Scholar 

  2. Chambers, T. J., Revell, P. A., Fuller, K., and Athanasou, N. (1984) Resorption of bone by isolated rabbit osteoclasts. J. Cell Sci. 66, 383–399.

    PubMed  CAS  Google Scholar 

  3. Hoebertz, A., Meghji, S., Burnstock, G., and Arnett, T. R. (2001) Extracellular ADP is a powerful stimulator of bone resorption: evidence for its signaling through the P2Y1 receptor. FASEB J. 15, 1139–1148.

    Article  PubMed  CAS  Google Scholar 

  4. Arnett, T. R. and Dempster, D. W. (1986) Effect of pH on bone resorption by rat osteoclasts in vitro. Endocrinology 119, 119–124.

    Article  PubMed  CAS  Google Scholar 

  5. Walsh, C. A., Beresford, J. N., Birch, M. A., Boothroyd, B., and Gallagher, J. A. (1991) Application of reflected light microscopy to identify and quantitate resorption by isolated osteoclasts. J. Bone Miner. Res. 6, 661–671.

    Article  PubMed  CAS  Google Scholar 

  6. Boyde, A. and Jones, S. J. (1991) Pitfalls in pit measurement. Calcif. Tissue Int. 49, 65–70.

    Article  PubMed  CAS  Google Scholar 

  7. Goldhaber, P. and Rabadjija, L. (1987) H+ stimulation of cell-mediated bone resorption in tissue culture. Am. J. Physiol. 253, E90–E98.

    PubMed  CAS  Google Scholar 

  8. Arnett, T. R. and Spowage, M. Modulation of resorptive activity of rat osteoclasts by small changes in extracellular pH near the physiological range. Bone 18, 277–279.

    Google Scholar 

  9. Morrison, M. S. and Arnett, T. R. (1997) Effect of extracellular pH on resorption pit formation by chick osteoclasts. J. Bone Miner. Res. 12, S290 (Abstr.).

    Article  Google Scholar 

  10. Morrison, M. S. and Arnett, T. R. (1998) pH effects on osteoclast formation and activation. Bone 22, 30S (Abstr.).

    Google Scholar 

  11. Hoebertz, A., Nesbitt, S. A., Horton, M. A., and Arnett, T. R. (1999) Acid activation of osteoclasts derived from human osteoclastoma. J. Bone Min. Res. 14, S1049 (Abstr.).

    Google Scholar 

  12. Meghji, S., Morrison, M. S., Henderson, B., and Arnett, T. R. (2001) pH-dependence of bone resorption: mouse calvarial osteoclasts are activated by acidosis. Am. J. Physiol. 280, E112–E119.

    CAS  Google Scholar 

  13. Murrills, R. J., Dempster, D. W., and Arnett, T. R. (1998) Isolation and culture of osteoclasts and osteoclast resorption assays, in Methods in Bone Biology (Arnett, T. R. and Henderson, B., eds.), Chapman and Hall, London, pp. 64–105.

    Google Scholar 

  14. Morrison, M. S., Turin, L., King, B. F., Burnstock, G., and Arnett, T.R. (1998) ATP is a potent stimulator of the activation and formation of rodent osteoclasts. J. Physiol. 511, 495–500.

    Article  PubMed  CAS  Google Scholar 

  15. Zanellato, N., Hoebertz, A., and Arnett, T. R. (2000) Low pH is a key requirement for osteoclast stimulation by RANKL. J. Bone Miner. Res. 15, S387 (Abstr.).

    Google Scholar 

  16. Bushinsky, D. A. (1987) Net calcium efflux from live bone during chronic metabolic, but not respiratory, acidosis. Am. J. Physiol. 256, F836–F842.

    Google Scholar 

  17. Arnett, T. R. and Dempster, D. W. (1987) A comparative study of disaggregated chick and rat osteoclasts in vitro: effects of calcitonin and prostaglandins. Endocrinology 120, 602–608.

    Article  PubMed  CAS  Google Scholar 

  18. Shibutani, T. and Heersche, J. N. (1993) Effect of medium pH on osteoclast activity and osteoclast formation in cultures of dispersed rabbit osteoclasts. J. Bone Miner. Res. 8, 331–336.

    Article  PubMed  CAS  Google Scholar 

  19. Lees, R. L., Sabharwal, V. K., and Heersche, J. N. (2001) Resorptive state and cell size influence intracellular pH regulation in rabbit osteoclasts cultured on collagen-hydroxyapatite films. Bone 28, 187–194.

    Article  PubMed  CAS  Google Scholar 

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© 2003 Humana Press Inc., Totowa, NJ

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Hoebertz, A., Arnett, T.R. (2003). Isolated Osteoclast Cultures. In: Helfrich, M.H., Ralston, S.H. (eds) Bone Research Protocols. Methods in Molecular Medicine, vol 80. Humana Press. https://doi.org/10.1385/1-59259-366-6:53

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  • DOI: https://doi.org/10.1385/1-59259-366-6:53

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-044-1

  • Online ISBN: 978-1-59259-366-8

  • eBook Packages: Springer Protocols

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