Fig. 1: Establishment of glandular patterned gastric epithelial model.

a A Polydimethylsiloxane (PDMS) device is employed to hold a patterned hydrogel as growth substrate for epithelial cells as published previously¹⁴. Gastric epithelial cells obtained from organoid cultures are seeded onto the hydrogel and let to grow until confluency. Apical and basal side media can be controlled independently. b Brightfield overview and detailed (extended depth of field, inset) images of fully established model 7 days after seeding. Representative images from > 20 independent experiments. c 3D reconstruction (left) and single planes (right) of confocal images of fixed sample stained for Nuclei (DAPI, blue), F-Actin (pink) and Mucus (UEA-FITC, green). Scale bar, 100 µm. d Confocal image of fluorescently stained cryosection of a 7-day old sample illustrating SOX9 and KI67-positive cells in the gland regions, including single-channel split-ups below. Representative images from three independent experiments. e Frequency of SOX9+ and KI67+ cells within the gland and surface regions, respectively. Each dot represents the quantification of the fraction of positive cells within one image with 1–3 glands, originating from three independent experiments. Bars represent mean and error bars SD values. Statistics were performed using a paired two-sided t-test. ****, p < 0.0001. Source data are provided as a Source Data file. f Confocal images of fluorescently stained cryosections of 7-day old samples illustrating cells positive for differentiation markers. Representative images from three independent experiments. Scale bars, 500 µm (b), 100 µm (c, d, f).