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. 2017 Aug 15;7:8290. doi: 10.1038/s41598-017-08197-2

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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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3-D culture of lacrimal gland spheres. Lacrimal gland epithelial cells from day 7 post-reopening in P2 formed similarly sized lacrimal spheres in suspension culture (A), and gradually branched out in about 2 weeks (days in culture, 5, (B); 10, (C)) to form mini gland-like structure in laminin gel. Immunostaining demonstrated K4 (green, D,E) positive, Lactoferrin (red, F,G) positive and AQP5 positive (green, F,G) cells in those structures. RT-qPCR showed the mRNA level of differentiated lacrimal gland cell-related genes in the gland-like structures (H–J). Blue for DAPI. Bar represents 100 μm.