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. 2019 Feb 25;9:2732. doi: 10.1038/s41598-019-39522-6

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© The Author(s) 2019

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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Co-stimulation system and fascicle-inspired engineered skeletal muscle tissue (eSMT). (a,b) The fascicle-inspired eSMT formed a cylindrical shape with length of 6 mm and diameter of approximately 75 μm (a). The tissues were stained using immunofluorescence technique to visualize the striations of α-actinin, which is a marker of differentiation and contractility (b, reproduced from reference 40 with permission from the Mary Ann Liebert, Inc., New Rochelle, NY). Scale bars represent 5 mm in (a) and 10 μm in (b). (c) Schematic of the experimental setup used to apply coordinated electric and mechanical stimulation to the eSMT. The co-stimulation system consists of electrodes for applying the electric potential, a cantilever wire moved by a servomotor, and a laser micrometre to monitor the displacement of the cantilever. The eSMT is pulled sideways with the cantilever to stretch eSMT to the desired strain. Contractile force was quantified by measuring deformation of the cantilever whose bending stiffness is known. (d,e) Cross-sectional images of unstimulated eSMTs stained for collagen IV (red) and actin (green) in longitudinal (d) and transverse (e) directions. Scale bars represent 10 μm.