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. 2018 Aug 17;8:12307. doi: 10.1038/s41598-018-29968-5

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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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Ectopic skeletal muscle regeneration. (A) Representative immunofluorescent images for MHC (red)/HLA (green) at 2 weeks after implantation. Double-immunostained MHC⁺/HLA⁺ myofibers in bioprinted constructs indicate newly formed skeletal muscle. (B) Numbers of MHC⁺ myofibers and (C) areas of MHC⁺ myofibers (n = 4, 3 random regions/sample). There is an increasing trend of skeletal muscle tissue formation with increasing cell density, but no significant difference between 30 and 50 × 10⁶ cells/ml at 1 week and 2 weeks after implantation (*P < 0.05 compared with 10 × 10⁶ cells/ml at 1 week, **P < 0.05 compared with 10 and 20 × 10⁶ cells/ml at 1 week, ^†P < 0.05 compared with 10 × 10⁶ cells/ml at 2 weeks, and ^††P < 0.05 compared with 10 and 20 × 10⁶ cells/ml at 2 weeks). (D) Numbers of HLA + myofibers and (E) areas of HLA⁺ myofibers (n = 4, 3 random regions/sample). There is no significant difference between 30 × 10⁶ cells/ml and 50 × 10⁶ cells/ml at 2 weeks after implantation.