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. 2020 Jun 10;10:9393. doi: 10.1038/s41598-020-66229-w

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© The Author(s) 2020

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 BC 2.5D organoids. Expression of urothelial cell markers, CK7, CK20, and UPK3A in 2.5D organoid cells at early (4–8) and late passages (15–20) (A, n = 4). Scale bar: 50 μm. Comparison of cell proliferation at day 1, 3, and 5 between 2.5D organoid at the early and late passage and their original 3D organoid. Cell proliferation was assayed by Prestoblue cell viability reagent and shown as fold increase relative to day 1 (B, n = 6). Results were expressed as mean ± S.E.M. *P < 0.05 vs. 3D organoid. Expression of a basal cell marker, CK5 in the 2.5D organoid cells and parental 3D organoids (C, n = 4). Scale bar: 50 μm. Plasticity of BC 2.5D organoids. Representative bright-field images of the reverted 3D organoids from 2.5D ones (D). Scale bar: 200 μm.