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. 2022 Aug 15;12:13818. doi: 10.1038/s41598-022-17548-7

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Wrinkles on the substrate are generated by contraction of the myotube. (a) The substrate used in this study and its schematics viewed from the side. The substrate is coated with a silicone gel (polydimethylsiloxane) on an 18 mm × 18 mm cover glass, and surface of the gel is treated by oxygen plasma. (b) Myotubes at day 5 of differentiation cultured on the substrate. Scale bar is 20 µm. (c) The 2-well chamber was connected to an electrical pulse generator with a pair of carbon electrodes for each well. The myotubes on the substrate were contracted with electrical pulses of 20 mA current, 20 ms duration, 980 ms interval, at 1 Hz (see Supplementary Movie 1). (d) The contraction of the myotubes in response to the electrical pulse generated wrinkles on the substrate. White arrows indicate wrinkles on the substrate.