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. 2020 Mar 9;10:4326. doi: 10.1038/s41598-020-61405-4

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© The Author(s) 2020

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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Tracheal reconstruction through co-transplantation of primary chondrocytes and iPSC-MSCs. (A) Newly formed cartilage (arrow, red color) is observed as assessed by safranin O staining. (B) Type II collagen-specific immune-dab staining shows a brown positive signal with a lacunae structure. (C) The defect site was also analyzed at week 4 for cartilage regeneration via reconstruction of micro-CT images. Yellow arrows indicate regenerative cartilage in the region of the tracheal scaffold implant. (D) Regenerative tracheal cartilage was calculated based on the quantity of white signal within the area of the region of interest (red⁺) in micro-CT images. (*P < 0.05). (E) Alcian blue staining shows re-epithelialization on the luminal surface in segmental tracheal defects. In particular, the Chd/iPSC-MSC group exhibited thick epithelial regeneration and cilia formation. (F) Quantitative analysis of the thickness of regenerative epithelium 4 weeks after surgery. The epithelial thicknesses were significantly increased in the Chd / iPSC-MSC groups than in the Chd groups (***P < 0.001).