phaidra.vetmeduni.ac.at o:2675 DOI 10.3390/organoids2030012 The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids 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. eng intestinal organoid; redox biology; IBD; glutathione; oxidative stress; ROS; redox imaging 2024-03-05T09:16:20.377Z LOMv1.0 Author BEGIN:VCARD VERSION:3.0 N:Csukovich;Georg; FN:Georg Csukovich X-ORCID:https://orcid.org/0000-0001-5615-666X END:VCARD BEGIN:VCARD VERSION:3.0 N:Burgener;Iwan Anton; FN:Iwan Anton Burgener X-ORCID:https://orcid.org/0000-0002-8516-0040 END:VCARD BEGIN:VCARD VERSION:3.0 N:Pratscher;Barbara; FN:Barbara Pratscher X-ORCID:https://orcid.org/0000-0001-5291-1365 END:VCARD BEGIN:VCARD VERSION:3.0 N:Huainig;Janina; FN:Janina Huainig END:VCARD BEGIN:VCARD VERSION:3.0 N:Troester;Selina; FN:Selina Troester X-ORCID:https://orcid.org/0000-0001-6268-4442 END:VCARD https://w3id.org/kim/hcrt/scheme https://w3id.org/kim/hcrt/text Textdokument Text LOMv1.0 yes https://creativecommons.org/licenses/by/4.0 application/pdf 447854 https://phaidra.vetmeduni.ac.at/o:2675 https://phaidra.vetmeduni.ac.at/api/object/o:2675/thumbnail LOMv1.0 discipline