Characterization of bovine and ovine basal-out and apical-out ileum organoids

Organoids are three-dimensional stem cell-derived structures that differentiate into multiple cell types. Their capacity to self-organize, coupled with the presence of diverse cell types, means that organoids resemble their organ of origin in architecture and function. Organoids from intestinal tiss...

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Main Authors: Ambre F. Chapuis, Tanith Harte, Daniel R. G. Price, Marc N. Faber, William M. Anderson, Barbara Shih, Jayne C. Hope, Jo Moore, David Smith
Format: Article
Language:English
Published: The Royal Society 2025-07-01
Series:Royal Society Open Science
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Online Access:https://royalsocietypublishing.org/doi/10.1098/rsos.250326
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author Ambre F. Chapuis
Tanith Harte
Daniel R. G. Price
Marc N. Faber
William M. Anderson
Barbara Shih
Jayne C. Hope
Jo Moore
David Smith
author_facet Ambre F. Chapuis
Tanith Harte
Daniel R. G. Price
Marc N. Faber
William M. Anderson
Barbara Shih
Jayne C. Hope
Jo Moore
David Smith
author_sort Ambre F. Chapuis
collection DOAJ
description Organoids are three-dimensional stem cell-derived structures that differentiate into multiple cell types. Their capacity to self-organize, coupled with the presence of diverse cell types, means that organoids resemble their organ of origin in architecture and function. Organoids from intestinal tissues have been extensively used as a three-dimensional model for in vitro studies of the gut. However, they typically self-organize with basal-out polarity when cultured in a three-dimensional extracellular matrix scaffold, presenting a hurdle for experiments that require access to the apical epithelial surface. Methods to invert the surface polarity of intestinal organoids have been reported, but little information exists on how this change of polarity impacts gene expression and cell populations present within the organoids. To address this knowledge gap, we modelled both polarity states in intestinal organoids from two different ruminant species. Apical-out organoids largely retained the same gene expression profile as basal-out organoids. Moreover, a combination of RNA-seq and immunohistochemistry analyses demonstrated the retention of specific markers of enterocytes, enteroendocrine, goblet and tuft cells present in organoids of both polarity states. This study presents a comprehensive validation of apical-out ileal organoids, providing supporting evidence for the utility of this model in experiments that require access to the apical surface.
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spelling doaj-art-46b5ba3b04694800a070287a8bf7a41c2025-08-20T03:09:35ZengThe Royal SocietyRoyal Society Open Science2054-57032025-07-0112710.1098/rsos.250326Characterization of bovine and ovine basal-out and apical-out ileum organoidsAmbre F. Chapuis0Tanith Harte1Daniel R. G. Price2Marc N. Faber3William M. Anderson4Barbara Shih5Jayne C. Hope6Jo Moore7David Smith8Moredun Research Institute, Penicuik, Scotland, UKMoredun Research Institute, Penicuik, Scotland, UKMoredun Research Institute, Penicuik, Scotland, UKMoredun Research Institute, Penicuik, Scotland, UKMoredun Research Institute, Penicuik, Scotland, UKThe University of Edinburgh The Roslin Institute, Edinburgh, Scotland, UKThe University of Edinburgh The Roslin Institute, Edinburgh, Scotland, UKMoredun Research Institute, Penicuik, Scotland, UKMoredun Research Institute, Penicuik, Scotland, UKOrganoids are three-dimensional stem cell-derived structures that differentiate into multiple cell types. Their capacity to self-organize, coupled with the presence of diverse cell types, means that organoids resemble their organ of origin in architecture and function. Organoids from intestinal tissues have been extensively used as a three-dimensional model for in vitro studies of the gut. However, they typically self-organize with basal-out polarity when cultured in a three-dimensional extracellular matrix scaffold, presenting a hurdle for experiments that require access to the apical epithelial surface. Methods to invert the surface polarity of intestinal organoids have been reported, but little information exists on how this change of polarity impacts gene expression and cell populations present within the organoids. To address this knowledge gap, we modelled both polarity states in intestinal organoids from two different ruminant species. Apical-out organoids largely retained the same gene expression profile as basal-out organoids. Moreover, a combination of RNA-seq and immunohistochemistry analyses demonstrated the retention of specific markers of enterocytes, enteroendocrine, goblet and tuft cells present in organoids of both polarity states. This study presents a comprehensive validation of apical-out ileal organoids, providing supporting evidence for the utility of this model in experiments that require access to the apical surface.https://royalsocietypublishing.org/doi/10.1098/rsos.250326apical-outbasal-outthree-dimensional organoidsin vitro culture systemsLGR5+ stem cellscrypts
spellingShingle Ambre F. Chapuis
Tanith Harte
Daniel R. G. Price
Marc N. Faber
William M. Anderson
Barbara Shih
Jayne C. Hope
Jo Moore
David Smith
Characterization of bovine and ovine basal-out and apical-out ileum organoids
Royal Society Open Science
apical-out
basal-out
three-dimensional organoids
in vitro culture systems
LGR5+ stem cells
crypts
title Characterization of bovine and ovine basal-out and apical-out ileum organoids
title_full Characterization of bovine and ovine basal-out and apical-out ileum organoids
title_fullStr Characterization of bovine and ovine basal-out and apical-out ileum organoids
title_full_unstemmed Characterization of bovine and ovine basal-out and apical-out ileum organoids
title_short Characterization of bovine and ovine basal-out and apical-out ileum organoids
title_sort characterization of bovine and ovine basal out and apical out ileum organoids
topic apical-out
basal-out
three-dimensional organoids
in vitro culture systems
LGR5+ stem cells
crypts
url https://royalsocietypublishing.org/doi/10.1098/rsos.250326
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