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. 2021 Feb 10;11:3492. doi: 10.1038/s41598-021-82178-4

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Histological, immunocytochemical, and FF-OCT images of multilayered cultivated cell sheets. (a) After the complete culture and formation of mCLESs, haematoxylin and eosin (H&E) staining of the cross-sectional cell product revealed multilayered cell products. (b) FF-OCT cross-sectional image of mCLESs. (a,b) Sections 1, 2, and 3 represent the basal cell layer at 5 µm, wing cell layer at 20 µm, and superficial squamous epithelial layer at 40 µm above the bottom of the culture plate, respectively. (c,e,g) En face images of whole-mount immunocytochemical staining of mCLESs obtained from in vitro confocal microscopy at Section 1 (g), Section 2 (e), and Section 3 (c). Red: Actin. Blue: Hoechst 33,258 for the nucleus. (d,f,h) FF-OCT en face images of mCLESs at Section 1 (h), Section 2 (f) and Section 3 (d). Scale bar = 20 µm. mCLESs = multilayer cultivated limbal epithelial sheets.