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Ensemble Analysis of Angiogenic Growth in Three-Dimensional Microfluidic Cell Cultures

Author(s)
Farahat, Waleed A.; Wood, Levi Benjamin; Zervantonakis, Ioannis; Schor, Alisha R.; Ong, Lee Ling; Neal, Devin M.; Kamm, Roger Dale; Asada, Harry; ... Show more Show less
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Abstract
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.
Date issued
2012-05
URI
http://hdl.handle.net/1721.1/71755
Department
Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Mechanical Engineering
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
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.
Version: Final published version
ISSN
1932-6203
1932-6203

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