Abstract
We describe an equibiaxial cell stretcher and hybrid, elastic membrane platform designed for dynamic imaging of cells on substrates with physiological stiffness undergoing cyclic stretch. Studies enabled by this device revealed that both substrate stiffness and cyclic stretch coordinately protect pulmonary endothelial monolayers against thrombin-induced disruption. The fluorescence imaging possible with the designed hybrid membranes further revealed similarities and differences in actin and cell dynamics during monolayer recovery. The improved live-cell imaging capabilities of this platform, when used in conjunction with fluorescent probes, will have broad applications for investigations of the impact of biochemical stimuli and mechanotransduction mechanisms on mechanically perturbed tissues.
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Acknowledgments
We thank Brian Helmke (University of Virginia) for sharing their stretcher design. This work was funded by NSF CMMI 14-62739, NIH 5R01 GMS 097443 to D.E.L., and by Widiger and Ullyot Graduate Research Fellowships to A.D.
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Associate Editor Konstantinos Konstantopoulos oversaw the review of this article.
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Dan, A., Huang, R.B. & Leckband, D.E. Dynamic Imaging Reveals Coordinate Effects of Cyclic Stretch and Substrate Stiffness on Endothelial Integrity. Ann Biomed Eng 44, 3655–3667 (2016). https://doi.org/10.1007/s10439-016-1677-4
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DOI: https://doi.org/10.1007/s10439-016-1677-4