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. 2018 Apr 23;8:6400. doi: 10.1038/s41598-018-24694-4

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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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Macroscopic and microscopic evaluation of iAM and dAM. Cryopreserved iAM mounted on the culture insert showed sub-transparent appearance (A). Phase contrast image of the iAM whole mount showed uniformly arranged and well maintained epithelial cells (B). H&E (C) and DAPI staining (D) of iAM crosssection showed organized epithelial layer. Transparent dAM after 0.02% EDTA treatment and electric toothbrush scraping (E). Phase contrast image of the dAM whole mount showed few epithelial cells (F). H&E staining (G) and DAPI staining (H) of dAM crosssection confirmed the presence of an intact stroma without the epithelial layer. Immunostaining of collagen IV (I,L), collagen VII (J,M) and Laminin 5 (K,N) showed all these basement membrane components were well preserved after amniotic membrane epithelial denudation. Bars represent 500 μm in (B and F), and represent 100 μm in (C,D,G,H, I–N).