Investigating Intestinal Barrier Breakdown in Living Organoids

Marco Bardenbacher, Barbara Ruder, Natalie Britzen-Laurent, Elisabeth Naschberger, Christoph Becker, Ralph Palmisano, Michael Stürzl, Philipp Tripal

Research output: Contribution to journalOriginal Articlepeer-review

Abstract

Organoids and three-dimensional (3D) cell cultures allow the investigation of complex biological mechanisms and regulations in vitro, which previously was not possible in classical cell culture monolayers. Moreover, monolayer cell cultures are good in vitro model systems but do not represent the complex cellular differentiation processes and functions that rely on 3D structure. This has so far only been possible in animal experiments, which are laborious, time consuming, and hard to assess by optical techniques. Here we describe an assay to quantitatively determine the barrier integrity over time in living small intestinal mouse organoids. To validate our model, we applied interferon gamma (IFN-γ) as a positive control for barrier destruction and organoids derived from IFN-γ receptor 2 knock out mice as a negative control. The assay allowed us to determine the impact of IFN-γ on the intestinal barrier integrity and the IFN-γ induced degradation of the tight junction proteins claudin-2, -7, and -15. This assay could also be used to investigate the impact of chemical compounds, proteins, toxins, bacteria, or patient-derived probes on the intestinal barrier integrity.

Original languageEnglish
JournalJOURNAL OF VISUALIZED EXPERIMENTS
Issue number157
DOIs
Publication statusPublished - 26 Mar 2020
Externally publishedYes

Keywords

  • Animals
  • Humans
  • Image Processing, Computer-Assisted
  • Intestines/physiology
  • Mice
  • Models, Biological
  • Organoids/physiology
  • Permeability

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