Isolation, Purification, Generation, and Culture of Osteocytes

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

Research output: Book/ReportBook

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.

Original languageEnglish
PublisherHumana Press
Number of pages13
Volume1914
ISBN (Electronic)978-1-4939-8997-3
ISBN (Print)978-1-4939-8996-6
DOIs
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology (Clifton, N.J.)
PublisherSpringer Verlag
ISSN (Print)1064-3745

Fingerprint

Osteocytes
Osteoblasts
Cell Line
Bone Matrix
Skeleton
Phenotype
Gene Expression
Bone and Bones

Keywords

  • Cell purification
  • Osteoblast
  • Osteocyte

Cite this

Gooi, J. H., Chia, L. Y., Vrahnas, C., & Sims, N. A. (2019). Isolation, Purification, Generation, and Culture of Osteocytes. (Methods in Molecular Biology (Clifton, N.J.)). Humana Press. https://doi.org/10.1007/978-1-4939-8997-3_3
Gooi, Jonathan H. ; Chia, Ling Yeong ; Vrahnas, Christina ; Sims, Natalie A. / Isolation, Purification, Generation, and Culture of Osteocytes. Humana Press, 2019. 13 p. (Methods in Molecular Biology (Clifton, N.J.)).
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Gooi, JH, Chia, LY, Vrahnas, C & Sims, NA 2019, Isolation, Purification, Generation, and Culture of Osteocytes. Methods in Molecular Biology (Clifton, N.J.), vol. 1914, Humana Press. https://doi.org/10.1007/978-1-4939-8997-3_3

Isolation, Purification, Generation, and Culture of Osteocytes. / Gooi, Jonathan H.; Chia, Ling Yeong; Vrahnas, Christina; Sims, Natalie A.

Humana Press, 2019. 13 p. (Methods in Molecular Biology (Clifton, N.J.)).

Research output: Book/ReportBook

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N2 - 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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Gooi JH, Chia LY, Vrahnas C, Sims NA. Isolation, Purification, Generation, and Culture of Osteocytes. Humana Press, 2019. 13 p. (Methods in Molecular Biology (Clifton, N.J.)). https://doi.org/10.1007/978-1-4939-8997-3_3