A machine learning‑based image segmentation method to quantify in vitro osteoclast culture endpoints
Article
Davies, B.K., Hibbert, A.P., Roberts, S.J., Roberts, H., Tickner, J.C., Holdsworth, G., Arnett, T.R. and Orriss, I.R. 2023. A machine learning‑based image segmentation method to quantify in vitro osteoclast culture endpoints. Calcified Tissue International and Musculoskeletal Research. 113 (4), p. 437–448.
Type | Article |
---|---|
Title | A machine learning‑based image segmentation method to quantify in vitro osteoclast culture endpoints |
Authors | Davies, B.K., Hibbert, A.P., Roberts, S.J., Roberts, H., Tickner, J.C., Holdsworth, G., Arnett, T.R. and Orriss, I.R. |
Abstract | Quantification of in vitro osteoclast cultures (e.g. cell number) often relies on manual counting methods. These approaches are labour intensive, time consuming and result in substantial inter- and intra-user variability. This study aimed to develop and validate an automated workflow to robustly quantify in vitro osteoclast cultures. Using ilastik, a machine learning-based image analysis software, images of tartrate resistant acid phosphatase-stained mouse osteoclasts cultured on dentine discs were used to train the ilastik-based algorithm. Assessment of algorithm training showed that osteoclast numbers strongly correlated between manual- and automatically quantified values (r = 0.87). Osteoclasts were consistently faithfully segmented by the model when visually compared to the original reflective light images. The ability of this method to detect changes in osteoclast number in response to different treatments was validated using zoledronate, ticagrelor, and co-culture with MCF7 breast cancer cells. Manual and automated counting methods detected a 70% reduction (p < 0.05) in osteoclast number, when cultured with 10 nM zoledronate and a dose-dependent decrease with 1-10 μM ticagrelor (p < 0.05). Co-culture with MCF7 cells increased osteoclast number by ≥ 50% irrespective of quantification method. Overall, an automated image segmentation and analysis workflow, which consistently and sensitively identified in vitro osteoclasts, was developed. Advantages of this workflow are (1) significantly reduction in user variability of endpoint measurements (93%) and analysis time (80%); (2) detection of osteoclasts cultured on different substrates from different species; and (3) easy to use and freely available to use along with tutorial resources. |
Keywords | Formation; Ilastik; Machine learning; Osteoclast; Resorption |
Sustainable Development Goals | 3 Good health and well-being |
Middlesex University Theme | Health & Wellbeing |
Publisher | Springer |
Journal | Calcified Tissue International and Musculoskeletal Research |
ISSN | 0171-967X |
Electronic | 1432-0827 |
Publication dates | |
Online | 11 Aug 2023 |
31 Oct 2023 | |
Publication process dates | |
Accepted | 29 Jul 2023 |
Submitted | 06 Jun 2023 |
Deposited | 16 Aug 2023 |
Output status | Published |
Publisher's version | License |
PubMed ID | 37566229 |
Web of Science identifier | WOS:001046126700001 |
Language | English |
https://repository.mdx.ac.uk/item/9v792
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