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. 2021 Apr 21;12:247. doi: 10.1186/s13287-021-02133-y

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Generation of epithelial organoids from human lacrimal glands (LGs). A Scheme for the development and culture of lacrimal tissue-derived organoid. B Optimization of culture media following organoid formation and subculture. Representative images were obtained by light microscopy after culturing for 3 days (Magnification; 10×, Scale bar; 500 µm). C The growth of organoid cultured in optimized LG organoid medium (LGOM). Scale 100 μm. D A comparison of histological analysis between normal lacrimal tissue and formed LG organoid. H&E, Alcian blue, Masson’s trichrome, and PAS staining were performed for identifying the acini structure, mucosubstance secretion, connective tissue, and glycoprotein secretion, respectively (Magnification; 40×, Scale bar; 100 µm). E Confirmation of acinar cells (green; aquaporin 5; AQP5) and epithelial lineage (red; EpCAM) cells in lacrimal tissue (Magnification; 100×, Scale bar; 50 µm). F Organoid formation from EpCAM-positive cell fraction. Representative bright-field images (Magnification; 10×, Scale bar; 500 µm) were obtained after 4 days of cultivation and used for counting the formed organoid (right, graph, N = 4). G Ki67, AQP5, and vimentin (VIM) expression was detected in the organoids and observed using immunofluorescence (Magnification; 100×, Scale bar; 50 µm)