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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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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