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. 2017 Apr 10;7:777. doi: 10.1038/s41598-017-00914-1

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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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Characterization of the flat collagen sheet and the perfusion system for corneal tissue engineering. (A) Photograph of a flat collagen sheet. (B) Representative section of the collagen sheet. (C) Tensile stress-strain curve for each sample. (D) Cell density was calculated based on the total number of cells in each square millimetre. (E) Cells at different passages on TCPS and on collagen sheets were stained to detect SA-β-Gal activity. Scale bar: 100 μm. (F) The senescence level of the CECs was evaluated based on the ratio of SA-β-Gal-positive cells to total cells. (G) SEM images of the collagen sheet prepared under vitrification conditions and representative images of CECs on collagen sheets in static and perfusion culture. Differences with *P < 0.05 were considered statistically significant.