Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures
We demonstrate ensemble three-dimensional cell cultures and quantitative analysis of angiogenic growth from uniform endothelial monolayers. Our approach combines two key elements: a micro-fluidic assay that enables parallelized angiogenic growth instances subject to common extracellular conditions,...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Public Library of Science
2012
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| Online Access: | http://hdl.handle.net/1721.1/71755 https://orcid.org/0000-0002-9898-2377 https://orcid.org/0000-0003-3155-6223 https://orcid.org/0000-0002-7232-304X |
| _version_ | 1811091992500764672 |
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| author | Farahat, Waleed A. Wood, Levi Benjamin Zervantonakis, Ioannis Schor, Alisha R. Ong, Lee Ling Neal, Devin M. Kamm, Roger Dale Asada, Harry |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Farahat, Waleed A. Wood, Levi Benjamin Zervantonakis, Ioannis Schor, Alisha R. Ong, Lee Ling Neal, Devin M. Kamm, Roger Dale Asada, Harry |
| author_sort | Farahat, Waleed A. |
| collection | MIT |
| description | We demonstrate ensemble three-dimensional cell cultures and quantitative analysis of angiogenic growth from uniform endothelial monolayers. Our approach combines two key elements: a micro-fluidic assay that enables parallelized angiogenic growth instances subject to common extracellular conditions, and an automated image acquisition and processing scheme enabling high-throughput, unbiased quantification of angiogenic growth. Because of the increased throughput of the assay in comparison to existing three-dimensional morphogenic assays, statistical properties of angiogenic growth can be reliably estimated. We used the assay to evaluate the combined effects of vascular endothelial growth factor (VEGF) and the signaling lipid sphingoshine-1-phosphate (S1P). Our results show the importance of S1P in amplifying the angiogenic response in the presence of VEGF gradients. Furthermore, the application of S1P with VEGF gradients resulted in angiogenic sprouts with higher aspect ratio than S1P with background levels of VEGF, despite reduced total migratory activity. This implies a synergistic effect between the growth factors in promoting angiogenic activity. Finally, the variance in the computed angiogenic metrics (as measured by ensemble standard deviation) was found to increase linearly with the ensemble mean. This finding is consistent with stochastic agent-based mathematical models of angiogenesis that represent angiogenic growth as a series of independent stochastic cell-level decisions. |
| first_indexed | 2024-09-23T15:11:12Z |
| format | Article |
| id | mit-1721.1/71755 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:11:12Z |
| publishDate | 2012 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/717552022-10-02T01:13:18Z Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures Farahat, Waleed A. Wood, Levi Benjamin Zervantonakis, Ioannis Schor, Alisha R. Ong, Lee Ling Neal, Devin M. Kamm, Roger Dale Asada, Harry Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Mechanical Engineering Kamm, Roger Dale Farahat, Waleed A. Wood, Levi Benjamin Zervantonakis, Ioannis Schor, Alisha R. Neal, Devin M. Kamm, Roger Dale Asada, Harry We demonstrate ensemble three-dimensional cell cultures and quantitative analysis of angiogenic growth from uniform endothelial monolayers. Our approach combines two key elements: a micro-fluidic assay that enables parallelized angiogenic growth instances subject to common extracellular conditions, and an automated image acquisition and processing scheme enabling high-throughput, unbiased quantification of angiogenic growth. Because of the increased throughput of the assay in comparison to existing three-dimensional morphogenic assays, statistical properties of angiogenic growth can be reliably estimated. We used the assay to evaluate the combined effects of vascular endothelial growth factor (VEGF) and the signaling lipid sphingoshine-1-phosphate (S1P). Our results show the importance of S1P in amplifying the angiogenic response in the presence of VEGF gradients. Furthermore, the application of S1P with VEGF gradients resulted in angiogenic sprouts with higher aspect ratio than S1P with background levels of VEGF, despite reduced total migratory activity. This implies a synergistic effect between the growth factors in promoting angiogenic activity. Finally, the variance in the computed angiogenic metrics (as measured by ensemble standard deviation) was found to increase linearly with the ensemble mean. This finding is consistent with stochastic agent-based mathematical models of angiogenesis that represent angiogenic growth as a series of independent stochastic cell-level decisions. National Science Foundation (U.S.). Office of Emerging Frontiers in Research and Innovation (grant #0735997) 2012-07-23T17:38:50Z 2012-07-23T17:38:50Z 2012-05 2011-02 Article http://purl.org/eprint/type/JournalArticle 1932-6203 1932-6203 http://hdl.handle.net/1721.1/71755 Farahat, Waleed A. et al. “Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures.” Ed. Christophe Egles. PLoS ONE 7.5 (2012): e37333. https://orcid.org/0000-0002-9898-2377 https://orcid.org/0000-0003-3155-6223 https://orcid.org/0000-0002-7232-304X en_US http://dx.doi.org/10.1371/journal.pone.0037333 PLoS ONE Creative Commons Attribution http://creativecommons.org/licenses/by/2.5/ application/pdf Public Library of Science PLoS |
| spellingShingle | Farahat, Waleed A. Wood, Levi Benjamin Zervantonakis, Ioannis Schor, Alisha R. Ong, Lee Ling Neal, Devin M. Kamm, Roger Dale Asada, Harry Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures |
| title | Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures |
| title_full | Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures |
| title_fullStr | Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures |
| title_full_unstemmed | Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures |
| title_short | Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures |
| title_sort | ensemble analysis of angiogenic growth in three dimensional microfluidic cell cultures |
| url | http://hdl.handle.net/1721.1/71755 https://orcid.org/0000-0002-9898-2377 https://orcid.org/0000-0003-3155-6223 https://orcid.org/0000-0002-7232-304X |
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