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. 2017 Oct 20;7:13708. doi: 10.1038/s41598-017-14053-0

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© The Author(s) 2017

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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Light microscopic images of tissue sections of vascularized iPSC-CM 3D-tissues 28 days after implantation to rat infarcted heart. (a) Transverse section of rat heart implanted with the 3D-tissue with hematoxylin-eosin (H.E.) staining. The area encircled by the dotted line indicates the implanted tissue in the infarcted left ventricle (LV). (b) Immunostaining for cTnT (red) depicts cardiomyocytes both in the implant and host residual myocardium. Cellular nuclei are stained with DAPI. (c) Immunostaining for cTnT (red) and Cx43 (green) shows a fine meshwork of iPSC-CMs with substantially-ranged sarcomeres and many reaction products for Cx43 on them. Cellular nuclei are stained with DAPI. Inset: Higher magnification of the boxed area in c. Scale bar: 5 µm. (d) Immunostaining for human CD31 (red) and multi-species CD31 (green) shows many host-derived vessels (arrows) immunopositive only for multi-species CD31 and a few implant-derived vessels (arrowheads) immunopositive both for human and multi-species CD31 (yellow) within the implant. The dotted line indicates the border between the host and implant tissue. (e) Higher magnification of an implant-derived vessel (arrowhead) immunopositive both for human and multi-species CD31 (yellow). Inset: The H.E.-stained tissue section adjacent to e shows blood contents within the implant-derived vessels (arrowhead). Scale bar: 40 µm.