A simple microfluidic device to study cell-scale endothelial mechanotransduction
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Atherosclerosis is triggered by chronic inflammation of arterial endothelial cells (ECs). Because atherosclerosis develops preferentially in regions where blood flow is disturbed and where ECs have a cuboidal morphology, the interplay between EC shape and mechanotransduction events is of primary interest. In this work we present a simple microfluidic device to study relationships between cell shape and EC response to fluid shear stress. Adhesive micropatterns are used to non-invasively control EC elongation and orientation at both the monolayer and single cell levels. The micropatterned substrate is coupled to a microfluidic chamber that allows precise control of the flow field, high-resolution live-cell imaging during flow experiments, and in situ immunostaining. Using micro particle image velocimetry, we show that cells within the chamber alter the local flow field so that the shear stress on the cell surface is significantly higher than the wall shear stress in regions containing no cells. In response to flow, we observe the formation of lamellipodia in the downstream portion of the EC and cell retraction in the upstream portion. We quantify flow-induced calcium mobilization at the single cell level for cells cultured on unpatterned surfaces or on adhesive lines oriented either parallel or orthogonal to the flow. Finally, we demonstrate flow-induced intracellular calcium waves and show that the direction of propagation of these waves is determined by cell polarization rather than by the flow direction. The combined versatility and simplicity of this microfluidic device renders it very useful for studying relationships between EC shape and mechanosensitivity.
KeywordsAtherosclerosis Mechanobiology Shear stress Calcium signaling Micropatterns Microfluidic flow chamber
The authors thank Bertrand Levaché for introducing them to the double-sided tape microfabrication technique and Maria Isabella Gariboldi for her participation in micropatterning technique development. This work was supported in part by an endowment in cardiovascular cellular engineering from the AXA Research Fund. Julie Lafaurie-Janvore was funded by postdoctoral fellowships from the Fondation Lefoulon-Delalande and the AXA Research Fund. Elizabeth Antoine was funded by a Whitaker International Program postdoctoral fellowship. Sidney J. Perkins was supported by a summer research international student fellowship from École polytechnique and the Columbia University European Institute’s Fellowship Program.
- F. An, Q. Yueyang, X. Liu, R. Zhong, Y. Luo, Organ-on-a-Chip: new platform for biological analysis. Anal. Chem. Insights. 10, 39–45 (2015)Google Scholar
- K. A. Barbee, T. Mundel, R. Lal, P. F. Davies, Subcellular distribution of shear stress at the surface of flow-aligned and nonaligned endothelial monolayers. Am. J. Physiol. Heart Circ. Physiol. 268, H1765–H1772 (1995)Google Scholar
- Y. S. Chatzizisis, A. U. Coskun, M. Jonas, E. R. Edelman, C. L. Feldman, P. H. Stone, Role of endothelial shear stress in the natural history of coronary atherosclerosis and vascular remodeling: molecular, cellular, and vascular behavior. J. Am. Coll. Cardiol. 49(25), 2379–2393 (2007)CrossRefGoogle Scholar
- P. F. Davies, Flow-mediated endothelial mechanotransduction. Physiol. Rev. 75(3), 519–560 (1995)Google Scholar
- A. Edelstein, N. Amodaj, K. Hoover, R. Vale, N. Stuurman, Computer control of microscopes using μManager. Curr. Protoc. Mol. Biol. Chapter 14, 14.20.1–14.20.17 (2010)Google Scholar
- E. Fröhlich, G. Bonstingl, A. Höfler, C. Meindl, G. Leitinger, T. R. Pieber, E. Roblegg, Comparison of two in vitro systems to assess cellular effects of nanoparticles-containing aerosols. Toxicol. in Vitro. 27(1), 409–417 (2013)Google Scholar
- R. G. Parton, M. A. del Pozo, Caveolae as Plasma Membrane Sensors, Protectors and Organizers. Nat. Rev. Mol. Cell. Biol. 14(2) Nature Publishing Group, 98–112 (2013)Google Scholar
- W. J. Polacheck, R. Li, S. G. M. Uzel, R. D. Kamm, Microfluidic platforms for mechanobiology. Lab. Chip. 13(12), 2252–2267 (2013)Google Scholar
- J. Shemesh, I. Jalilian, A. Shi, G. H. Yeoh, T. M. L. Knothe, M. E. Warkiani, Flow-induced stress on adherent cells in microfluidic devices. Lab .Chip. Royal Society of Chemistry (2015). doi: 10.1039/C5LC00633C