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An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids

The process of sprouting angiogenesis can be measured in vitro using endothelial cells in sprouting assays such as the fibrin bead assay and the spheroid-based assay. While the technical aspects of these sprouting assays have been well-optimized, the analysis aspects have been limited to manual meth...

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Autores principales: Kannan, Pavitra, Schain, Martin, Lane, David P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9093605/
https://www.ncbi.nlm.nih.gov/pubmed/35571133
http://dx.doi.org/10.3389/fphar.2022.883083
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author Kannan, Pavitra
Schain, Martin
Lane, David P.
author_facet Kannan, Pavitra
Schain, Martin
Lane, David P.
author_sort Kannan, Pavitra
collection PubMed
description The process of sprouting angiogenesis can be measured in vitro using endothelial cells in sprouting assays such as the fibrin bead assay and the spheroid-based assay. While the technical aspects of these sprouting assays have been well-optimized, the analysis aspects have been limited to manual methods, which can be time-consuming and difficult to reproduce. Here, we developed an automated analysis tool called AQuTAS to quantify sprouting parameters from the spheroid-based sprouting assay. We trained and validated the algorithm on two subsets of data, and tested its sensitivity by measuring changes in sprouting parameters over a range of concentrations of pro- and antiangiogenic compounds. Our results demonstrate that the algorithm detects known differences in sprouting parameters in endothelial spheroids treated with pro- and antiangiogenic compounds. Moreover, it is sensitive to biological changes that are ≥40%. Among the five quantified parameters, cumulative sprout length is likely the most discriminative parameter for measuring differences in sprouting behavior because it had the highest effect size (>1.5 Cohen’s d). In summary, we have generated an automated tool that quantifies sprouting parameters from the spheroid-based assay in a reproducible and sensitive manner.
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spelling pubmed-90936052022-05-12 An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids Kannan, Pavitra Schain, Martin Lane, David P. Front Pharmacol Pharmacology The process of sprouting angiogenesis can be measured in vitro using endothelial cells in sprouting assays such as the fibrin bead assay and the spheroid-based assay. While the technical aspects of these sprouting assays have been well-optimized, the analysis aspects have been limited to manual methods, which can be time-consuming and difficult to reproduce. Here, we developed an automated analysis tool called AQuTAS to quantify sprouting parameters from the spheroid-based sprouting assay. We trained and validated the algorithm on two subsets of data, and tested its sensitivity by measuring changes in sprouting parameters over a range of concentrations of pro- and antiangiogenic compounds. Our results demonstrate that the algorithm detects known differences in sprouting parameters in endothelial spheroids treated with pro- and antiangiogenic compounds. Moreover, it is sensitive to biological changes that are ≥40%. Among the five quantified parameters, cumulative sprout length is likely the most discriminative parameter for measuring differences in sprouting behavior because it had the highest effect size (>1.5 Cohen’s d). In summary, we have generated an automated tool that quantifies sprouting parameters from the spheroid-based assay in a reproducible and sensitive manner. Frontiers Media S.A. 2022-04-27 /pmc/articles/PMC9093605/ /pubmed/35571133 http://dx.doi.org/10.3389/fphar.2022.883083 Text en Copyright © 2022 Kannan, Schain and Lane. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Kannan, Pavitra
Schain, Martin
Lane, David P.
An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids
title An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids
title_full An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids
title_fullStr An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids
title_full_unstemmed An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids
title_short An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids
title_sort automated quantification tool for angiogenic sprouting from endothelial spheroids
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9093605/
https://www.ncbi.nlm.nih.gov/pubmed/35571133
http://dx.doi.org/10.3389/fphar.2022.883083
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