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. 2017 Jun 20;8:15613. doi: 10.1038/ncomms15613

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Copyright © 2017, The Author(s)

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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(a) Perfusion of bioconstructs sustains MuSC viability in vitro. Bioconstructs were reconstituted with Luc⁺ MuSCs and Luc⁻ MRCs and then cultured for 24 h. Bioluminescence was measured at different time points, as indicated (n=6). (b) Model of the bioreactor designed to perfuse cultured bioconstructs. An autoclavable plexiglass chamber, capable of holding, in parallel, five independent tubing lines, is shown (top panel). These lines were connected to an external pump. Each line contained one tubular culture chamber capable of hosting one bioconstruct (bottom panel). (c) Representative bioluminescence and fluorescence images of bioconstructs before and after transplantation. Bioconstructs were generated with Luc⁺/GFP⁺ MuSCs and Luc⁻/GFP⁻ MRCs, cultured under either perfused or static conditions, and imaged non-invasively before transplantation (Pre Op, left panels). Bioconstructs that had been imaged previously in vitro were imaged non-invasively in vivo immediately following transplantation (Post Op, right panels). (d) Quantitative results comparing bioluminescence of perfused and static bioconstructs at periodic intervals following transplantation into VML injuries (n=4). (e) (Left panels) Representative H&E-stained cross-sections of muscles 10 days after receiving perfused or static bioconstruct transplantation to treat VML injuries. Bioconstructs (BC) are shown adjacent to unablated muscle (UM) (scale bar=500 μm). (Right panels) The regions outlined by the yellow boxes in the left panel are magnified here (scale bar=200 μm Data are±s.e.m. For statistical analysis, t-tests were used. *P<0.05; **P<0.001; ***P<0.0001.