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Bone Research Protocols pp 39–51Cite as

Isolation, Purification, Generation, and Culture of Osteocytes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1914))

Abstract

Osteocytes reside within bone matrix and produce both paracrine and endocrine factors that influence the skeleton and other tissues. Despite their abundance and physiological importance, osteocytes have been difficult to study in vitro because they are difficult to extract and purify, and do not retain their phenotype in standard culture conditions. However, new techniques for this purpose are emerging. This chapter will describe three methods we use to study osteocytes: (1) isolating and purifying primary osteocytes from murine bone, with and without hematopoietic-lineage depletion, (2) differentiating cultured osteoblasts (or osteoblast cell lines) until they reach a stage of osteocytic gene expression, and (3) using the Ocy454 osteocyte-like cell line.

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Acknowledgment

The authors acknowledge the support of the National Health and Medical Research Council (Australia) (NHMRC) Project Grants 100242 and 100978. NAS is supported by an NHMRC Senior Research Fellowship. St Vincent’s Institute is also supported by State Government of Victoria’s Operational Infrastructure Support program.

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Authors and Affiliations

  1. Department of Medicine, St. Vincent’s Hospital Melbourne, The University of Melbourne, Melbourne, VIC, Australia

    Jonathan H. Gooi, Ling Yeong Chia, Christina Vrahnas & Natalie A. Sims

  2. St. Vincent’s Institute of Medical Research, Melbourne, VIC, Australia

    Ling Yeong Chia, Christina Vrahnas & Natalie A. Sims

  3. Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Clayton, VIC, Australia

    Ling Yeong Chia

  4. MRC Protein Phosphorylation & Ubiquitylation Unit, University of Dundee, Sir James Black Centre, Dundee, UK

    Christina Vrahnas

Authors
  1. Jonathan H. Gooi
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  2. Ling Yeong Chia
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  3. Christina Vrahnas
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  4. Natalie A. Sims
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Corresponding author

Correspondence to Natalie A. Sims .

Editor information

Editors and Affiliations

  1. Department of Oncology and Metabolism, University of Sheffield, Sheffield, UK

    Aymen I. Idris

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Gooi, J.H., Chia, L.Y., Vrahnas, C., Sims, N.A. (2019). Isolation, Purification, Generation, and Culture of Osteocytes. In: Idris, A. (eds) Bone Research Protocols. Methods in Molecular Biology, vol 1914. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8997-3_3

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  • DOI: https://doi.org/10.1007/978-1-4939-8997-3_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8996-6

  • Online ISBN: 978-1-4939-8997-3

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