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. 2018 Feb 13;8:2871. doi: 10.1038/s41598-018-21201-7

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© The Author(s) 2018

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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The primary human Intestine Chip exhibits multi-lineage differentiation. (a) Graph showing the fold change in the levels of transcripts from epithelial cells cultured in the Intestine Chip and assessed at day 12 relative to the transcript levels in the undifferentiated state assessed at the day 8 (the medium perfused through the apical channel was switched from EM to DM on day 8). Genes analyzed include sucrose-isomaltase (SI) and alkaline phosphatase (ALPI) that are specific for absorptive enterocytes; mucin 2 (MUC2) and mucin 5AC (MUC5AC) for goblet cells; chromogranin A (CHGA) and synaptophysin (SYP) for enteroendocrine cells; lysozyme (LYZ) for Paneth cells; and leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5) and polycomb complex protein BMI-1 (BMI1) for stem cells. Values are presented as mean ±SEM from 3 independent experiments involving Intestine Chips generated from organoids isolated from 3 different donors (ns, not significant; *p ≤ 0.05; ***p ≤ 0.001 by Student’s t-test). (b) A schematic cross-sectional representation of the 3D intestinal epithelial tissue architecture developed on chip (top) and confocal immunofluorescence micrographs (bottom) showing vertical cross-sections of the differentiated epithelium in Intestine Chip stained for lysozyme (Lyz, green), mucin 2 (Muc2, magenta), chromogranin A (ChgA, yellow) and villin (green). Cell nuclei were counterstained with DAPI (blue).