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. 2021 Mar 8;11:5401. doi: 10.1038/s41598-021-84774-w

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© Crown 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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Morphology and characteristics of rabbit small intestinal organoids. (a) L-WRN-conditioned medium supported the growth of spheroids from duodenum, jejunum and ileum spheroids from a laboratory rabbit, and duodenum spheroids from a wild rabbit. Scale bars = 500 μm. (b) Proliferating rabbit duodenal spheroids and (c) organoids were imaged either unstained (brightfield), after staining with hematoxylin and eosin (H&E) or after immunostaining with a CD44-specific (green) antibody; nuclei were counterstained with DAPI (blue). (d) Expression of stem cell-related genes (AXIN2 and LGR5) in differentiated organoids. Data are presented as fold change (2^−ΔΔCt) in gene expression from undifferentiated spheroids, calculated from three individual cell culture wells with three technical RT-qPCR replicates each. Error bars represent standard errors of the mean. Student’s t-test was performed to assess the statistical significance; only statistically significant differences are shown (*p < 0.05). (e) Representative images of organoids after four days of culturing in differentiation medium; the images show representative organoids with either a cystic (left panel) or multilobular morphology (right panel). Scale bars = 100 μm. b–e Used duodenal spheroids/organoids from a single laboratory rabbit.