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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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Decellularisation protocols. (a) Diagram showing the process used to prepare rat hind limbs for decellularisation. The femoral artery is cannulated and the hind limb dissected; decellularised reagents where delivered through the vessel tree by using a pump directly connected to the cannula. Three different decellularisation methods were performed, and called LatB, DET and SDS. (b) Diagram showing steps used to perform LatB, DET and SDS decellularisation. (c) Quantification of DNA content in freshly isolated rat skeletal muscle and LatB-, DET- and SDS-decellularised muscles. Data are shown as mean ± s.e.m of three independent replicates; **P < 0.01, one-way ANOVA and Tukey’s multiple comparison test; no significant differences were observed among the decellularised samples; n = 6–10, each group. (d) Macroscopic and microscopic (H&E and scanning electron microscopy, SEM) evaluation of fresh rat muscles and LatB-, DET- and SDS-decellularised samples. For H&E analysis, cross-sections were analysed. Scale bars, macroscopic: 1 cm; H&E: 100 µm; SEM: 10 µm.