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High throughput, quantitative analysis of human osteoclast differentiation and activity
Natalie A Diepenhorst1, Cameron J Nowell1, Patricia Rueda1
1Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC 3052, Australia.
Analytical Biochemistry
|December 19, 2016
Summary
A new imaging method quantifies osteoclast activity at the single-cell level using tartrate-resistant acid phosphatase (TRAP). This technique reveals TRAP activity correlates with osteoclast maturity, aiding osteoporosis drug development.
Area of Science:
- Cell Biology
- Biochemistry
- Pharmacology
Background:
- Osteoclasts are crucial for bone remodeling and are therapeutic targets for osteoporosis.
- Current methods assess osteoclast activity at the population level, overlooking single-cell maturation.
- Tartrate-resistant acid phosphatase (TRAP) is a key marker for osteoclast activity.
Purpose of the Study:
- To develop a quantitative, single-cell method for measuring osteoclast TRAP activity.
- To correlate osteoclast TRAP activity with its nucleation state (maturity).
- To enable high-throughput screening for novel osteoporosis therapeutics.
Main Methods:
- Developed a multi-labelled imaging assay using a fluorescent TRAP substrate.
- Employed automated image analysis with open-source software for high-throughput assessment.
- Investigated the impact of receptor activator of nuclear factor kappa-B ligand (RANK-L) on osteoclast activity.
Main Results:
- Established a method for precise, single-cell quantification of osteoclast TRAP activity.
- Demonstrated a direct correlation between TRAP activity and osteoclast nuclei number (maturity).
- Showcased the method's utility in evaluating RANK-L effects on osteoclast maturation.
Conclusions:
- The developed imaging method offers a sensitive, high-throughput approach to assess osteoclast function.
- This technique advances the understanding of osteoclast maturation and activity.
- It provides a valuable tool for discovering and developing osteoporosis treatments.
