High throughput, quantitative analysis of human osteoclast differentiation and activity

Natalie Diepenhorst, Cameron Nowell, Patricia Rueda, Kim Henriksen, Tracie Pierce, Anna Cook, Philippe Pastoureau, Massimo Sabatini, William Charman, Arthur Christopoulos, Roger Summers, Patrick Sexton, Christopher Langmead

Research output: Contribution to journalArticleResearchpeer-review

2 Citations (Scopus)

Abstract

Osteoclasts are multinuclear cells that degrade bone under both physiological and pathophysiological conditions. Osteoclasts are therefore a major target of osteoporosis therapeutics aimed at preserving bone. Consequently, analytical methods for osteoclast activity are useful for the development of novel biomarkers and/or pharmacological agents for the treatment of osteoporosis. The nucleation state of an osteoclast is indicative of its maturation and activity. To date, activity is routinely measured at the population level with only approximate consideration of the nucleation state (an ‘osteoclast population’ is typically defined as cells with ≥3 nuclei). Using a fluorescent substrate for tartrate-resistant acid phosphatase (TRAP), a routinely used marker of osteoclast activity, we developed a multi-labelled imaging method for quantitative measurement of osteoclast TRAP activity at the single cell level. Automated image analysis enables interrogation of large osteoclast populations in a high throughput manner using open source software. Using this methodology, we investigated the effects of receptor activator of nuclear factor kappa-B ligand (RANK-L) on osteoclast maturation and activity and demonstrated that TRAP activity directly correlates with osteoclast maturity (i.e. nuclei number). This method can be applied to high throughput screening of osteoclast-targeting compounds to determine changes in maturation and activity.

Original languageEnglish
Pages (from-to)51-56
Number of pages6
JournalAnalytical Biochemistry
Volume519
DOIs
Publication statusPublished - 15 Feb 2017

Keywords

  • Bone resorption
  • High throughput
  • ImageJ
  • Osteoclast
  • Quantitative image analysis
  • Tartrate-resistant acid phosphatase (TRAP)

Cite this

@article{0dbb56d424bf41588eb88becc09ab070,
title = "High throughput, quantitative analysis of human osteoclast differentiation and activity",
abstract = "Osteoclasts are multinuclear cells that degrade bone under both physiological and pathophysiological conditions. Osteoclasts are therefore a major target of osteoporosis therapeutics aimed at preserving bone. Consequently, analytical methods for osteoclast activity are useful for the development of novel biomarkers and/or pharmacological agents for the treatment of osteoporosis. The nucleation state of an osteoclast is indicative of its maturation and activity. To date, activity is routinely measured at the population level with only approximate consideration of the nucleation state (an ‘osteoclast population’ is typically defined as cells with ≥3 nuclei). Using a fluorescent substrate for tartrate-resistant acid phosphatase (TRAP), a routinely used marker of osteoclast activity, we developed a multi-labelled imaging method for quantitative measurement of osteoclast TRAP activity at the single cell level. Automated image analysis enables interrogation of large osteoclast populations in a high throughput manner using open source software. Using this methodology, we investigated the effects of receptor activator of nuclear factor kappa-B ligand (RANK-L) on osteoclast maturation and activity and demonstrated that TRAP activity directly correlates with osteoclast maturity (i.e. nuclei number). This method can be applied to high throughput screening of osteoclast-targeting compounds to determine changes in maturation and activity.",
keywords = "Bone resorption, High throughput, ImageJ, Osteoclast, Quantitative image analysis, Tartrate-resistant acid phosphatase (TRAP)",
author = "Natalie Diepenhorst and Cameron Nowell and Patricia Rueda and Kim Henriksen and Tracie Pierce and Anna Cook and Philippe Pastoureau and Massimo Sabatini and William Charman and Arthur Christopoulos and Roger Summers and Patrick Sexton and Christopher Langmead",
year = "2017",
month = "2",
day = "15",
doi = "10.1016/j.ab.2016.12.010",
language = "English",
volume = "519",
pages = "51--56",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Elsevier",

}

High throughput, quantitative analysis of human osteoclast differentiation and activity. / Diepenhorst, Natalie; Nowell, Cameron; Rueda, Patricia; Henriksen, Kim; Pierce, Tracie; Cook, Anna; Pastoureau, Philippe; Sabatini, Massimo; Charman, William; Christopoulos, Arthur; Summers, Roger; Sexton, Patrick; Langmead, Christopher.

In: Analytical Biochemistry, Vol. 519, 15.02.2017, p. 51-56.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - High throughput, quantitative analysis of human osteoclast differentiation and activity

AU - Diepenhorst, Natalie

AU - Nowell, Cameron

AU - Rueda, Patricia

AU - Henriksen, Kim

AU - Pierce, Tracie

AU - Cook, Anna

AU - Pastoureau, Philippe

AU - Sabatini, Massimo

AU - Charman, William

AU - Christopoulos, Arthur

AU - Summers, Roger

AU - Sexton, Patrick

AU - Langmead, Christopher

PY - 2017/2/15

Y1 - 2017/2/15

N2 - Osteoclasts are multinuclear cells that degrade bone under both physiological and pathophysiological conditions. Osteoclasts are therefore a major target of osteoporosis therapeutics aimed at preserving bone. Consequently, analytical methods for osteoclast activity are useful for the development of novel biomarkers and/or pharmacological agents for the treatment of osteoporosis. The nucleation state of an osteoclast is indicative of its maturation and activity. To date, activity is routinely measured at the population level with only approximate consideration of the nucleation state (an ‘osteoclast population’ is typically defined as cells with ≥3 nuclei). Using a fluorescent substrate for tartrate-resistant acid phosphatase (TRAP), a routinely used marker of osteoclast activity, we developed a multi-labelled imaging method for quantitative measurement of osteoclast TRAP activity at the single cell level. Automated image analysis enables interrogation of large osteoclast populations in a high throughput manner using open source software. Using this methodology, we investigated the effects of receptor activator of nuclear factor kappa-B ligand (RANK-L) on osteoclast maturation and activity and demonstrated that TRAP activity directly correlates with osteoclast maturity (i.e. nuclei number). This method can be applied to high throughput screening of osteoclast-targeting compounds to determine changes in maturation and activity.

AB - Osteoclasts are multinuclear cells that degrade bone under both physiological and pathophysiological conditions. Osteoclasts are therefore a major target of osteoporosis therapeutics aimed at preserving bone. Consequently, analytical methods for osteoclast activity are useful for the development of novel biomarkers and/or pharmacological agents for the treatment of osteoporosis. The nucleation state of an osteoclast is indicative of its maturation and activity. To date, activity is routinely measured at the population level with only approximate consideration of the nucleation state (an ‘osteoclast population’ is typically defined as cells with ≥3 nuclei). Using a fluorescent substrate for tartrate-resistant acid phosphatase (TRAP), a routinely used marker of osteoclast activity, we developed a multi-labelled imaging method for quantitative measurement of osteoclast TRAP activity at the single cell level. Automated image analysis enables interrogation of large osteoclast populations in a high throughput manner using open source software. Using this methodology, we investigated the effects of receptor activator of nuclear factor kappa-B ligand (RANK-L) on osteoclast maturation and activity and demonstrated that TRAP activity directly correlates with osteoclast maturity (i.e. nuclei number). This method can be applied to high throughput screening of osteoclast-targeting compounds to determine changes in maturation and activity.

KW - Bone resorption

KW - High throughput

KW - ImageJ

KW - Osteoclast

KW - Quantitative image analysis

KW - Tartrate-resistant acid phosphatase (TRAP)

UR - http://www.scopus.com/inward/record.url?scp=85007201355&partnerID=8YFLogxK

U2 - 10.1016/j.ab.2016.12.010

DO - 10.1016/j.ab.2016.12.010

M3 - Article

VL - 519

SP - 51

EP - 56

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

ER -