Isolation, Purification, Generation, and Culture of Osteocytes

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

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

3 Citations (Scopus)


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.

Original languageEnglish
Title of host publicationBone Research Protocols
EditorsAymen I Idris
Place of PublicationNew York NY USA
PublisherHumana Press
Number of pages13
ISBN (Electronic)9781493989973
ISBN (Print)9781493989966
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Cell purification
  • Osteoblast
  • Osteocyte

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