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. 2021 Nov 16;12:6610. doi: 10.1038/s41467-021-26762-2

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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 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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Fig. 2 — a Selective withdrawal of gastric organoid cytokines from control (CTRL) complete medium, in CS 23 (mid-week 8) organoid line split at single cells at passage 7. Scale bar 400 μm. b Bright field images of representative organoid line for each of the 3 stages, showing the formation of spherical organoids within 7 days starting from single cells. Scale bar 100 μm. c Immunofluorescence panel showing ezrin (EZR) in red, f-actin (F-ACT) in cyan, mucin 5AC (MUC5AC) in green, and nuclei in blue (Hoechst). Scale bars 25 μm. d Cumulative cell counts during days of culture. Mean ± SD (n = 4 biological replicates for early and late fetal, n = 3 for pediatric organoids). e Single-nucleotide polymorphism (SNP) array for pediatric organoids. Representative image of the chromosome viewer and allele frequency. f Real-time PCR analysis of early fetal, late fetal, and pediatric gastric organoids. Relative fold change to GAPDH. Mean ± SEM (n = 3). Black circles on the bar charts represent single biological replicates. Ordinary one-way ANOVA.