Isolation of osteocytes from human trabecular bone

Matthew Prideaux, Christine Schutz, Asiri R. Wijenayaka, David M. Findlay, David G. Campbell, Lucian B. Solomon, Gerald J. Atkins

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Osteocytes are essential regulators of bone homeostasis. However, they are difficult to study due to their location within the bone mineralised matrix. Although several techniques have been published for the isolation of osteocytes from mouse bone, no such technique has been described for human osteocytes. We have therefore developed a protocol for the isolation of osteocytes from human trabecular bone samples acquired during surgery. The cells were digested from the bone matrix by sequential collagenase and ethylenediaminetetraacetic acid (EDTA) digestions and the cells from later digests displayed characteristic dendritic osteocyte morphology when cultured ex vivo. Furthermore, the cells expressed characteristic osteocyte marker genes, such as E11, dentin matrix protein 1 (DMP1), SOST, matrix extracellular phosphoglycoprotein (MEPE) and phosphate regulating endopeptidase homologue, X-linked (PHEX). In addition, genes associated with osteocyte perilacunar remodelling, including matrix metallopeptidase-13 (MMP13), cathepsin K (CTSK) and carbonic anhydrase 2 (CAR2) were expressed. The cells also responded to parathyroid hormone (PTH) by downregulating SOST mRNA expression and to 1α,25-dihydroxyvitamin D3 (1,25D) by upregulating fibroblast growth factor 23 (FGF23) mRNA expression. Therefore, the cells behave in a similar manner to osteocytes in vivo. These cells represent an important tool in enhancing current knowledge in human osteocyte biology.

Original languageEnglish (US)
Pages (from-to)64-72
Number of pages9
JournalBone
Volume88
DOIs
StatePublished - Jul 1 2016
Externally publishedYes

Fingerprint

Osteocytes
Bone Matrix
PHEX Phosphate Regulating Neutral Endopeptidase
Cathepsin K
Cancellous Bone
Bone and Bones
Messenger RNA
Carbonic Anhydrases
Calcitriol
Metalloproteases
Collagenases
Dentin
Parathyroid Hormone
Edetic Acid
Genes
Extracellular Matrix
Digestion
Homeostasis
Down-Regulation

Keywords

  • Cell morphology
  • Gene expression
  • Osteocyte
  • Trabecular bone

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Histology
  • Physiology

Cite this

Prideaux, M., Schutz, C., Wijenayaka, A. R., Findlay, D. M., Campbell, D. G., Solomon, L. B., & Atkins, G. J. (2016). Isolation of osteocytes from human trabecular bone. Bone, 88, 64-72. https://doi.org/10.1016/j.bone.2016.04.017

Isolation of osteocytes from human trabecular bone. / Prideaux, Matthew; Schutz, Christine; Wijenayaka, Asiri R.; Findlay, David M.; Campbell, David G.; Solomon, Lucian B.; Atkins, Gerald J.

In: Bone, Vol. 88, 01.07.2016, p. 64-72.

Research output: Contribution to journalArticle

Prideaux, M, Schutz, C, Wijenayaka, AR, Findlay, DM, Campbell, DG, Solomon, LB & Atkins, GJ 2016, 'Isolation of osteocytes from human trabecular bone', Bone, vol. 88, pp. 64-72. https://doi.org/10.1016/j.bone.2016.04.017
Prideaux M, Schutz C, Wijenayaka AR, Findlay DM, Campbell DG, Solomon LB et al. Isolation of osteocytes from human trabecular bone. Bone. 2016 Jul 1;88:64-72. https://doi.org/10.1016/j.bone.2016.04.017
Prideaux, Matthew ; Schutz, Christine ; Wijenayaka, Asiri R. ; Findlay, David M. ; Campbell, David G. ; Solomon, Lucian B. ; Atkins, Gerald J. / Isolation of osteocytes from human trabecular bone. In: Bone. 2016 ; Vol. 88. pp. 64-72.
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