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. 2018 Apr 12;8:5898. doi: 10.1038/s41598-018-24273-7

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Cell morphology and quantification of cell alignment. (a) Cell morphology over time. (a) day 0, cells are randomly distributed immediately after seeding; (b) day 1, cells attach and elongate under static conditions; (c) day 3, cells start to become aligned under flow for one day; (d) day 4, most of the cells are aligned under flow for two days. Arrows indicate the direction of pulsatile flow in the microfluidic channels. (e) F-actin staining of PAEC in static conditions and (f) under flow. If not specified scale bar represents 100 µm. (b,c) Quantification of cell alignment to the x-axis of the microfluidic channels by immunofluorescence staining for the cytoskeleton protein F-actin and CD31, respectively. On the left panel, column graphs of the average cell angle in degrees to the x-axis are shown under static and pulsatile flow conditions (mean values ± SD, p-value: * < 0.05, ** < 0.01). Representative immunofluorescence images are shown on the right panel (a-b). Arrows show the flow direction. Scale bar represents 50 µm.