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. 2024 Dec 2;14:29921. doi: 10.1038/s41598-024-80946-6

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© The Author(s) 2024

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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Fig. 2 — Construction of gut MPS/Fluid3D-X. Cells were cultured for 11 days, and phase-contrast images of gut MPS/Fluid3D-X were captured using an APX100 microscope (EVIDENT CORPORATION). Bright-field images of (a) liquid-liquid interface and (b) air-liquid interface cultures. Bar: 2 mm. (c) Cultured cells under ALI conditions (day 12) were observed using an optical coherence tomography (OCT) imaging system (CELL3 IMAGER ESTIER, SCREEN Holdings Co., LTD.). Top left: Bright field image of the observation area. Blue line indicates the scan direction. Top right: X-Z section image of yellow line (top left). The images were acquired at a 3 μm pitch across a 1 mm² area. Bottom left: 3D dome structure obtained from the top. The image was generated utilizing Image J software (National Institutes of Health, Bethesda, MD, USA) Bottom right: 3D dome structure taken from the side. Scale bars: 250 μm. (d) Time-dependent expression of mRNAs (days 8, 11, and 13) was confirmed by probes that detect transporters, P450 enzymes, intestinal epithelial markers, and stem cell markers. The absolute value of each copy number was divided by that of EpCAM1. * Copy numbers were less than the lower limit of the calibration curve. Each bar represents the mean value (n = 2).