Abstract
We used collagen microtissues (~200 µm) as a base to grow endothelial monolayers and assess their barrier function in vitro. Using this ECM-based platform, we confirmed endothelial cells adhere and form characteristic intercellular junctions. Using optical sectioning, we measured the endothelial permeability to be 2.76 ± 0.61 x10-8 cm/s. To improve the speed of this functional assessment, we used wide-field imaging to assess endothelial barrier function in a high-throughput manner (n>=471). With this rapid imaging method, we assessed the effect of adding the endothelial coating to the collagen microtissues, finding the change in barrier function to be statistically significant (p<0.01).
| Original language | English (US) |
|---|---|
| Title of host publication | 21st International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2017 |
| Publisher | Chemical and Biological Microsystems Society |
| Pages | 997-998 |
| Number of pages | 2 |
| ISBN (Electronic) | 9780692941836 |
| State | Published - Jan 1 2020 |
| Event | 21st International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2017 - Savannah, United States Duration: Oct 22 2017 → Oct 26 2017 |
Publication series
| Name | 21st International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2017 |
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Conference
| Conference | 21st International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2017 |
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| Country/Territory | United States |
| City | Savannah |
| Period | 10/22/17 → 10/26/17 |
Bibliographical note
Funding Information:We have shown that behaviors seen in other endothelial mimics (confluence of the monolayer, and behavior of the semi-permeable barrier) are also observed in our smaller-scale platform. Additionally, we have shown, for the first time to the authors’ knowledge, high-throughput assessment (n>=471) of endothelial function on natural ECM, supporting robust statistical analysis. Miniaturization of tissues also minimizes cell number requirements, making this platform amenable to precious cell and ECM sources. We will continue exploring the potential of this platform, measuring the effect of inflammatory cytokines (e.g. TNF-α), and investigating cell-cell interactions by encapsulating cells (e.g. stromal cells) in the ECM of the constructs. ACKNOWLEDGEMENTS We thank the Minnesota Nanofabrication Center, the American Heart Association (AHA 13SDG6450000), the National Heart, Lung, and Blood Institute (NHLBI R21 HL132256), the National Institute of Environmental Health Sciences (NIEHS R21 ES027622), the National Science Foundation (GRFP Grant 00039202). REFERENCES [1]. DA Chistiakov et al. Endothelial Barrier and Its Abnormalities in Cardiovascular Disease. Frontiers in Phys-iology. 2015. [2]. P Totoson et al. Endothelial Dysfunction in Rheumatoid Arthritis: Mechanistic Insights and Correlation with CirculatingMarkersofSystemicInflammation.MillerF,ed.PLoS ONE. 2016. [3] PW Kazakoff, et al. "An in vitro model for endothelial permeability: assessment of monolayer integrity. "In VitroCellular&DevelopmentalBiology-Animal. 1995. [4]. M-E Brett*, AL. Crampton*, and DK. Wood. "Rapid generation of collagen-based microtissues to study cell– matrix interactions."Technology. 2016. CONTACT * D.K. Wood; phone: (612) 624-1438; [email protected]
Publisher Copyright:
© 17CBMS-0001.
Keywords
- Collagen
- Endothelial permeability
- Microfluidic
- Microtissue