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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2022.883083/full |
| _version_ | 1818009618726518784 |
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| author | Pavitra Kannan Martin Schain David P. Lane |
| author_facet | Pavitra Kannan Martin Schain David P. Lane |
| author_sort | Pavitra Kannan |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-14T05:45:01Z |
| format | Article |
| id | doaj.art-9401b0eb564e439896a8c78f63d6e2f8 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-04-14T05:45:01Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-9401b0eb564e439896a8c78f63d6e2f82022-12-22T02:09:19ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-04-011310.3389/fphar.2022.883083883083An Automated Quantification Tool for Angiogenic Sprouting From Endothelial SpheroidsPavitra Kannan0Martin Schain1David P. Lane2Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, SwedenAntaros Medical AB, Mölndal, SwedenDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, SwedenThe 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.https://www.frontiersin.org/articles/10.3389/fphar.2022.883083/fullsprouting angiogenesisautomated quantificationspheroidsin vitro angiogenesis assaysprouting parameters |
| spellingShingle | Pavitra Kannan Martin Schain David P. Lane An Automated Quantification Tool for Angiogenic Sprouting From Endothelial Spheroids Frontiers in Pharmacology sprouting angiogenesis automated quantification spheroids in vitro angiogenesis assay sprouting parameters |
| 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 | sprouting angiogenesis automated quantification spheroids in vitro angiogenesis assay sprouting parameters |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2022.883083/full |
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