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. 2017 Jul 26;7:6640. doi: 10.1038/s41598-017-06794-9

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© The Author(s) 2017

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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Change of MSC Shape as a Function of Cell Orientation, Stretch Direction and Substrate. Here are three schematic representations of the changes that 3 hypothetical MSCs undergo as a result of stretch and initial orientation (A). Cell nuclei are in blue and cell outlines are in brown color and star-like to visualize the exaggerated changes in shape parallel and perpendicular to the direction of the applied stretch, which is left-to-right, horizontally. With parallel cell alignment (see the first row with the left most cell pointing to 0 degrees), an MSC becomes longer in the direction of stretch while simultaneously contracting perpendicular cell extensions. This change increases the length and decreases both circularity and roundness. With perpendicular cell alignment (see the second row with the left most cell pointing to 90 degrees), a single MSC first contracts in the perpendicular direction and extends along the direction of stretch, eventually looking like it simply reoriented. This change produces a rounder and more circular cell with a decreased length before additional stretch returns its shape to an elongated morphology. With an oblique cell alignment (see the third row with the left most cell pointing to 45 degrees), the elongated MSC is stretched and it contracts protrusions away from 90 degrees and elongates in the direction of stretch. Mathematically, this follows the perpendicular cell but to an exaggerated extent. Collectively, the morphological changes of these 3 hypothetical MSCs highlight how stretching differently aligned MSCs can change their resulting morphology. The next panels exhibit the differences in alignment and shape induced by seeding on different substrates (B). MSC seeded on collagen coated silicone aligned perpendicularly to the direction of stretch while MSC seeded at the same concentration on a collagen sheet aligned parallel to the direction of stretch. When the cells were embedded in a 3D hydrogel of collagen and stretched, the cells aligned parallel to the direction of stretch.