title: Osteocyte isolation and culture methods.
abstract: The aim of this paper is to present several popular methods for in vitro culture of osteocytes and osteocyte cell lines. Osteocytes are located extremely suitably within the calcified bone matrix to sense mechanical signals, and are equipped with a multitude of molecular features that allow mechanosensing. However, osteocytes are more than specialized mechanosensing cells. Several signaling molecules are preferentially produced by osteocytes, and osteocytes hold a tight reign over osteoblast and osteoclast formation and activity, but also have a role as endocrine cell, communicating with muscles or organs as remote as the kidneys. In order to facilitate further research into this fascinating cell type, three protocols will be provided in this paper. The first protocol will be on the culture of mouse (early) osteocyte cell lines, the second on the isolation and culture of primary mouse bone cells, and the third on the culture of fully embedded human osteocytes within their own three-dimensional bone matrix.
authors.affiliation: Department of Human Metabolism, The Mellanby Centre for Bone Research, The Medical School, The University of Sheffield , Sheffield, UK.
authors.forename: Karan M
authors.identifier: 0000-0001-9909-6409
authors.initials: KM
authors.lastname: Shah
authors.affiliation: Department of Biology, Winthrop University , Rock Hill, SC, USA.
authors.forename: Matt M
authors.initials: MM
authors.lastname: Stern
authors.affiliation: Department Mechanical Engineering, School of Computing and Engineering, Mechanical Engineering, University of Missouri-Kansas City, Kansas City, MO, USA; Engineering Systems Inc., Charlotte, NC, USA.
authors.forename: Amber R
authors.initials: AR
authors.lastname: Stern
authors.affiliation: Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands; Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, China.
authors.forename: Janak L
authors.initials: JL
authors.lastname: Pathak
authors.affiliation: Department of Clinical Chemistry, VU University Medical Center, MOVE Research Institute Amsterdam , Amsterdam, The Netherlands.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bravenboer
authors.affiliation: Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, MOVE Research Institute Amsterdam , Amsterdam, The Netherlands.
authors.forename: Astrid D
authors.initials: AD
authors.lastname: Bakker