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

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
Title of host publicationBone Research Protocols
EditorsAymen I Idris
Place of PublicationNew York NY USA
PublisherHumana Press
Chapter3
Pages39-51
Number of pages13
Volume1914
Edition3
ISBN (Electronic)9781493989973
ISBN (Print)9781493989966
DOIs
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Number3
Volume1914
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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. In A. I. Idris (Ed.), Bone Research Protocols (3 ed., Vol. 1914, pp. 39-51). (Methods in Molecular Biology; Vol. 1914, No. 3). New York NY USA: 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. Bone Research Protocols. editor / Aymen I Idris. Vol. 1914 3. ed. New York NY USA : Humana Press, 2019. pp. 39-51 (Methods in Molecular Biology; 3).
@inbook{ae9e6e15331c431c95d85dc97276e932,
title = "Isolation, Purification, Generation, and Culture of Osteocytes",
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.",
keywords = "Cell purification, Osteoblast, Osteocyte",
author = "Gooi, {Jonathan H.} and Chia, {Ling Yeong} and Christina Vrahnas and Sims, {Natalie A.}",
year = "2019",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-8997-3_3",
language = "English",
isbn = "9781493989966",
volume = "1914",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
number = "3",
pages = "39--51",
editor = "Idris, {Aymen I}",
booktitle = "Bone Research Protocols",
address = "United States",
edition = "3",

}

Gooi, JH, Chia, LY, Vrahnas, C & Sims, NA 2019, Isolation, Purification, Generation, and Culture of Osteocytes. in AI Idris (ed.), Bone Research Protocols. 3 edn, vol. 1914, Methods in Molecular Biology, no. 3, vol. 1914, Humana Press, New York NY USA, pp. 39-51. 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.

Bone Research Protocols. ed. / Aymen I Idris. Vol. 1914 3. ed. New York NY USA : Humana Press, 2019. p. 39-51 (Methods in Molecular Biology; Vol. 1914, No. 3).

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

TY - CHAP

T1 - Isolation, Purification, Generation, and Culture of Osteocytes

AU - Gooi, Jonathan H.

AU - Chia, Ling Yeong

AU - Vrahnas, Christina

AU - Sims, Natalie A.

PY - 2019/1/1

Y1 - 2019/1/1

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.

AB - 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.

KW - Cell purification

KW - Osteoblast

KW - Osteocyte

UR - http://www.scopus.com/inward/record.url?scp=85061125149&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-8997-3_3

DO - 10.1007/978-1-4939-8997-3_3

M3 - Chapter (Book)

SN - 9781493989966

VL - 1914

T3 - Methods in Molecular Biology

SP - 39

EP - 51

BT - Bone Research Protocols

A2 - Idris, Aymen I

PB - Humana Press

CY - New York NY USA

ER -

Gooi JH, Chia LY, Vrahnas C, Sims NA. Isolation, Purification, Generation, and Culture of Osteocytes. In Idris AI, editor, Bone Research Protocols. 3 ed. Vol. 1914. New York NY USA: Humana Press. 2019. p. 39-51. (Methods in Molecular Biology; 3). https://doi.org/10.1007/978-1-4939-8997-3_3