Organoids

9 pages, 1369 KiB

Open AccessCommunication

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

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

Organoids 2023, 2(3), 156-164; https://doi.org/10.3390/organoids2030012 - 03 Sep 2023

Cited by 1 | Viewed by 1347

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. Full article

(This article belongs to the Special Issue Intestinal Organoid)

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16 pages, 1461 KiB

Open AccessReview

Incorporating Immune Cells into Organoid Models: Essential for Studying Human Disease

by Ania Bogoslowski, Meilin An and Josef M. Penninger

Organoids 2023, 2(3), 140-155; https://doi.org/10.3390/organoids2030011 - 12 Aug 2023

Cited by 2 | Viewed by 3634

Abstract

Organoid-based research has made significant discoveries and contributions to our understanding of human organ function in both health and disease. To continue making progress, it is crucial to acknowledge the crucial role of the immune system in all organs. Various immune cells, such as macrophages, T cells, and neutrophils, are resident in almost all human tissues and play essential roles in organ homeostasis, function, and disease. Using diverse methods, researchers have begun integrating immune cells into organoid models, leading to more physiologically relevant models that better represent various aspects of human disease. These methods range from immune cell injection to co-culture and tissue expansion with existing immune cells. Immune cells can be sourced from mature patients or generated from stem cells as immature immune cells. The successful incorporation of immune cells into organoids will enhance our understanding of organ function and provide a more accurate approximation of human disease. Full article

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17 pages, 1681 KiB

Open AccessReview

Organoid Models and Next-Generation Sequencing for Bone Marrow and Related Disorders

by Magdalena Rausch, Neelam Iqbal, Shelly Pathak, Heather E. Owston and Payal Ganguly

Organoids 2023, 2(3), 123-139; https://doi.org/10.3390/organoids2030010 - 01 Jul 2023

Cited by 3 | Viewed by 2599

Abstract

Challenges to the musculoskeletal system negatively impact the quality of life of people suffering from them, leading to pain, a decline in mobility, genetic alterations, and potential disorders. The bone marrow (BM) forms an integral part of the musculoskeletal system responsible for erythropoiesis and optimal survival of the various immune and stem cells within the BM. However, due to its dynamic and complex three-dimensional (3D) structure, replicating the BM physiologically in traditional two-dimensional (2D) cell culture settings is often challenging, giving rise to the need for 3D in vitro models to better dissect the BM and its regeneration. Several researchers globally have been investigating various approaches to define an appropriate 3D model for their research. Organoids are novel preclinical models that provide a 3D platform for several tissues and have been analysed using next-generation sequencing (NGS) to identify new molecular pathways at the genetic level. The 3D in vitro models and organoids are increasingly considered important platforms for precision medicine. This review outlines the current knowledge of organoid and 3D in vitro models for the BM. We also discuss different types of 3D models which may be more adaptable for the BM. Finally, we critically review the NGS techniques used for such models and the future combination of these techniques. Full article

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Organoids, Volume 2, Issue 3 (September 2023) – 3 articles

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