Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis
Inflammatory Bowel Diseases (IBDs) are characterized by chronic intestinal inflammation and fibrosis, the latter being the predominant denominator for long-term complications. Epithelial and mesenchymal 2D cultures are highly utilized in vitro models for the preclinical evaluation of anti-inflammato...
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Format: | Article |
Language: | English |
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Wiley
2021-01-01
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Series: | Stem Cells International |
Online Access: | http://dx.doi.org/10.1155/2021/9929461 |
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author | Leonidas Kandilogiannakis Eirini Filidou Ioannis Drygiannakis Gesthimani Tarapatzi Stylianos Didaskalou Maria Koffa Konstantinos Arvanitidis Giorgos Bamias Vassilis Valatas Vasilis Paspaliaris George Kolios |
author_facet | Leonidas Kandilogiannakis Eirini Filidou Ioannis Drygiannakis Gesthimani Tarapatzi Stylianos Didaskalou Maria Koffa Konstantinos Arvanitidis Giorgos Bamias Vassilis Valatas Vasilis Paspaliaris George Kolios |
author_sort | Leonidas Kandilogiannakis |
collection | DOAJ |
description | Inflammatory Bowel Diseases (IBDs) are characterized by chronic intestinal inflammation and fibrosis, the latter being the predominant denominator for long-term complications. Epithelial and mesenchymal 2D cultures are highly utilized in vitro models for the preclinical evaluation of anti-inflammatory and antifibrotic therapies. More recently, human intestinal organoids (HIOs), a new 3D in vitro model derived from pluripotent stem cells, have the advantage to closely resemble the architecture of the intestinal mucosa. However, the appropriate timing for the study of inflammatory and fibrotic responses, during HIO development, has not been adequately investigated. We developed HIOs from the human embryonic stem cell line, H1, and examined the expression of mesenchymal markers during their maturation process. We also investigated the effect of inflammatory stimuli on the expression of fibrotic and immunological mediators. Serial evaluation of the expression of mesenchymal and extracellular matrix (ECM) markers revealed that HIOs have an adequately developed mesenchymal component, which gradually declines through culture passages. Specifically, CD90, collagen type I, collagen type III, and fibronectin were highly expressed in early passages but gradually diminished in late passages. The proinflammatory cytokines IL-1α and TNF-α induced the mRNA expression of fibronectin, collagen types I and III, tissue factor (TF), and alpha-smooth muscle actin (α-SMA) primarily in early passages. Similarly, HIOs elicited strong mRNA and protein mesenchymal (CXCL10) and epithelial (CXCL1, CCL2, CXCL8, and CCL20) chemokine responses in early but not late passages. In contrast, the epithelial tight junction components, CLDN1 and JAMA, responded to inflammatory stimulation independently of the culture passage. Our findings indicate that this HIO model contains a functional mesenchymal component, during early passages, and underline the significance of the mesenchymal cells’ fitness in inflammatory and fibrotic responses. Therefore, we propose that this model is suitable for the study of epithelial-mesenchymal interactions in early passages when the mesenchymal component is active. |
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institution | Kabale University |
issn | 1687-966X 1687-9678 |
language | English |
publishDate | 2021-01-01 |
publisher | Wiley |
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spelling | doaj-art-cc3dd1974ffd4b44800bfd09bfeb89852025-02-03T01:05:32ZengWileyStem Cells International1687-966X1687-96782021-01-01202110.1155/2021/99294619929461Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and FibrosisLeonidas Kandilogiannakis0Eirini Filidou1Ioannis Drygiannakis2Gesthimani Tarapatzi3Stylianos Didaskalou4Maria Koffa5Konstantinos Arvanitidis6Giorgos Bamias7Vassilis Valatas8Vasilis Paspaliaris9George Kolios10Laboratory of Pharmacology, Faculty of Medicine, Democritus University of Thrace, Alexandroupolis, GreeceLaboratory of Pharmacology, Faculty of Medicine, Democritus University of Thrace, Alexandroupolis, GreeceGastroenterology and Hepatology Research Laboratory, Medical School, University of Crete, Heraklion, GreeceLaboratory of Pharmacology, Faculty of Medicine, Democritus University of Thrace, Alexandroupolis, GreeceLaboratory of Molecular Cell Biology, Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis, GreeceLaboratory of Molecular Cell Biology, Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis, GreeceLaboratory of Pharmacology, Faculty of Medicine, Democritus University of Thrace, Alexandroupolis, GreeceGI-Unit, Third Department of Internal Medicine, National & Kapodistrian University of Athens, Sotiria Hospital, Athens, GreeceLaboratory of Pharmacology, Faculty of Medicine, Democritus University of Thrace, Alexandroupolis, GreeceTithon Biotech Inc., San Diego, USALaboratory of Pharmacology, Faculty of Medicine, Democritus University of Thrace, Alexandroupolis, GreeceInflammatory Bowel Diseases (IBDs) are characterized by chronic intestinal inflammation and fibrosis, the latter being the predominant denominator for long-term complications. Epithelial and mesenchymal 2D cultures are highly utilized in vitro models for the preclinical evaluation of anti-inflammatory and antifibrotic therapies. More recently, human intestinal organoids (HIOs), a new 3D in vitro model derived from pluripotent stem cells, have the advantage to closely resemble the architecture of the intestinal mucosa. However, the appropriate timing for the study of inflammatory and fibrotic responses, during HIO development, has not been adequately investigated. We developed HIOs from the human embryonic stem cell line, H1, and examined the expression of mesenchymal markers during their maturation process. We also investigated the effect of inflammatory stimuli on the expression of fibrotic and immunological mediators. Serial evaluation of the expression of mesenchymal and extracellular matrix (ECM) markers revealed that HIOs have an adequately developed mesenchymal component, which gradually declines through culture passages. Specifically, CD90, collagen type I, collagen type III, and fibronectin were highly expressed in early passages but gradually diminished in late passages. The proinflammatory cytokines IL-1α and TNF-α induced the mRNA expression of fibronectin, collagen types I and III, tissue factor (TF), and alpha-smooth muscle actin (α-SMA) primarily in early passages. Similarly, HIOs elicited strong mRNA and protein mesenchymal (CXCL10) and epithelial (CXCL1, CCL2, CXCL8, and CCL20) chemokine responses in early but not late passages. In contrast, the epithelial tight junction components, CLDN1 and JAMA, responded to inflammatory stimulation independently of the culture passage. Our findings indicate that this HIO model contains a functional mesenchymal component, during early passages, and underline the significance of the mesenchymal cells’ fitness in inflammatory and fibrotic responses. Therefore, we propose that this model is suitable for the study of epithelial-mesenchymal interactions in early passages when the mesenchymal component is active.http://dx.doi.org/10.1155/2021/9929461 |
spellingShingle | Leonidas Kandilogiannakis Eirini Filidou Ioannis Drygiannakis Gesthimani Tarapatzi Stylianos Didaskalou Maria Koffa Konstantinos Arvanitidis Giorgos Bamias Vassilis Valatas Vasilis Paspaliaris George Kolios Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis Stem Cells International |
title | Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis |
title_full | Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis |
title_fullStr | Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis |
title_full_unstemmed | Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis |
title_short | Development of a Human Intestinal Organoid Model for In Vitro Studies on Gut Inflammation and Fibrosis |
title_sort | development of a human intestinal organoid model for in vitro studies on gut inflammation and fibrosis |
url | http://dx.doi.org/10.1155/2021/9929461 |
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