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. 2018 May 30;8:8398. doi: 10.1038/s41598-018-26371-y

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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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Isolation and seeding of anucleate myofibres. (a) Single myofibres were isolated from fresh or decellularised rat EDL digested in collagenase I and collected under the microscope. (b) Yield of fibres per EDL, ***p < 0.001 vs Fresh, t-test. (c) Scanning electron microscopy of single fresh and anucleate myofibres from LatB- and SDS-decellularised EDL. Scale bar: first row 100 µm, second row 10 µm, third row 1 µm. (d) Murine MuSCs were seeded on single myofibres and fresh EDL fibres were cultured to activate resident MuSCs. (e) Cellular migration speed along single fibres measured 24 hours post-seeding; ***p < 0.001, ANOVA. (f) Representative images of GFP+ MuSCs migrating on LatB-scaffold derived anucleate fibres, scale bar: 10 µm. (g) Immunofluorescence for MyoD and Pax7 of wild-type MuSCs seeded on LatB and SDS anucleate myofibres after 72 hours of culture, scale bar: 100 µm. (h) Subpopulations of cells according to positivity for Pax7 and MyoD after 72 hours of culture with statistical analysis showing significance in respect to control (CTRL). *p < 0.05; **p < 0.01; ***p < 0.001 vs CTRL, t-test.