Zeitschriftenaufsatz | 2023 Open Access

The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids.

Autor:in

Csukovich, Georg; Huainig, Janina; Troester, Selina; Pratscher, Barbara; Burgener, Iwan Anton

Publikationen als Autor:in / Herausgeber:in der Vetmeduni

Pratscher, Barbara; Tröster, Selina; Csukovich, Georg; Burgener, Iwan

Journal

Organoids

Dokumententyp

Short Communication

CC Lizenz

CC-BY

Open Access Type

Gold

ISSN/eISSN

2674-1172 - 2674-1172

DOI

10.3390/organoids2030012

Repository Phaidra

Projekt

Identification of cell populations in canine intestinal organoids and primary epithelium for modelling of acute and chronic gastrointestinal diseases.

Weitere Details

Band

2

Startseite

156

Letzte Seite

164

Nummer

3

We evaluated the redox status, precisely glutathione levels, which have a major impact in cellular detoxification and antioxidant defence in IBD-derived and healthy intestinal organoids. Therefore, we wanted to explore the differences in terms of their redox balance and mitochondrial fitness. To this end, we introduced a Grx1-roGFP2 construct into the organoids by lentiviral transduction before performing a stress assay by treating the organoids with hydrogen peroxide and examined the GSH/GSSG ratio using confocal imaging. Using ratio imaging, we could detect statistically significant differences between healthy and IBD-derived samples. To gain more insight, we also performed a GSH/GSSG assay, which directly measured glutathione levels. This analysis revealed that both organoid lines had higher levels of oxidized glutathione due to the stress treatment demonstrated by a lower GSH/GSSG ratio compared to the untreated control. Nevertheless, the results showed no significant difference between healthy and IBD-derived organoids. We further challenged organoids with hydrogen peroxide after incubation with MitoTracker® to see if mitochondrial fitness might be different in IBD-derived organoids. However, these results were also very comparable. In summary, our preliminary findings indicate that both organoid lines demonstrate a well-functioning system in terms of analysis but show no clear difference between healthy and IBD-derived samples.

VetImaging

Canine Organoids for Gastrointestinal Disease Modelling.

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We evaluated the redox status, precisely glutathione levels, which have a major impact in cellular detoxification and antioxidant defence in IBD-derived and healthy intestinal organoids. Therefore, we wanted to explore the differences in terms of their redox balance and mitochondrial fitness. To this end, we introduced a Grx1-roGFP2 construct into the organoids by lentiviral transduction before performing a stress assay by treating the organoids with hydrogen peroxide and examined the GSH/GSSG ratio using confocal imaging. Using ratio imaging, we could detect statistically significant differences between healthy and IBD-derived samples. To gain more insight, we also performed a GSH/GSSG assay, which directly measured glutathione levels. This analysis revealed that both organoid lines had higher levels of oxidized glutathione due to the stress treatment demonstrated by a lower GSH/GSSG ratio compared to the untreated control. Nevertheless, the results showed no significant difference between healthy and IBD-derived organoids. We further challenged organoids with hydrogen peroxide after incubation with MitoTracker® to see if mitochondrial fitness might be different in IBD-derived organoids. However, these results were also very comparable. In summary, our preliminary findings indicate that both organoid lines demonstrate a well-functioning system in terms of analysis but show no clear difference between healthy and IBD-derived samples.

VetImaging

Canine Organoids for Gastrointestinal Disease Modelling.

Download
RDF/XML-Format
JSON-LD-Format
Turtle-Format
N3-Format