A permeability assay for mouse intestinal organoids

Stijn Aaron den Daas; Ugo Soffientini; Shilpa Chokshi; Gautam Mehta

doi:10.1016/j.xpro.2022.101365

A permeability assay for mouse intestinal organoids

STAR Protoc. 2022 May 1;3(2):101365. doi: 10.1016/j.xpro.2022.101365. eCollection 2022 Jun 17.

Authors

Stijn Aaron den Daas^{1

2}, Ugo Soffientini^{1

2}, Shilpa Chokshi^{1

2}, Gautam Mehta^{1

2

3}

Affiliations

¹ Roger Williams Institute of Hepatology, Foundation for Liver Research, London SE5 9NT, UK.
² School of Immunology and Microbial Diseases, Faculty of Life Sciences and Medicine, King's College London, London WC2R 2LS, UK.
³ Institute for Liver and Digestive Health, Royal Free Campus, University College London, London NW3 2PF, UK.

Abstract

Here, we describe an assay for intestinal permeability in mouse intestinal organoids, although this may also be adapted for other species. Propidium iodide (PI) does not penetrate intact biological membranes and thus cannot enter the lumen of intact organoids. Passage of PI within the lumen can be induced by tight junction disruption or epithelial cell death. This technique measures PI-stained extruded dead cells within the organoid lumen to analyze the effect of insults, toxins, or treatments on intestinal organoid permeability.

Keywords: Health Sciences; Model Organisms; Organoids.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Intestinal Mucosa*
Intestines
Mice
Organoids*
Permeability
Tight Junctions / metabolism