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. 2018 Mar 7;8:4123. doi: 10.1038/s41598-018-22401-x

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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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Metabolic activity and seeding of porcine aMSCs on polyurethane graft. (A) After 7 days of incubation, biopsies from the cellularized scaffold demonstrated predominantly viable cells (green, calcein) with small, isolated patches of dead cells (red, EtBr). (B) Cryosections of punch biopsies demonstrated many nuclei (red, EtBr) on the superficial aspect of the scaffold. (C) Medium glucose and lactate concentrations from the bioreactors fresh medium on Day 0, and conditioned medium on days 2, 5, and 7 (n = 8 bioreactors, *p < 0.05, Steel-Dwass, all pairs). (D) DNA concentrations extracted from the cellularized scaffold increased with time (n = 3, *p < 0.05, one-way ANOVA). (E,F) Immunocytochemistry showed that cells attached to the scaffold expressed the cytoskeletal protein, nestin (red; DAPI, blue, E) and alpha smooth muscle actin (green, aSMA; DAPI, blue, F). (G) Concentrations of VEGFA, GMCSF, IL1RA, IL6 and IL8 proteins in fresh and conditioned medium after 7/8 days from cellularized bioreactors. (n = 8 bioreactors), *p < 0.05 t-test. OA = Outer aspect. LA = Luminal aspect. Scale bars: A = 1 mm. B = 500 μm. E,F = 50 μm.