Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures

• Published in: Stem cells international Vol. 2023; pp. 3328655 - 13
• Main Authors: Xing, Chengfeng, Liang, Guili, Yu, Xin, Zhang, Anxing, Luo, Xiang, Liu, Yu, Tang, Zengli, Wu, Bian, Song, Zhengji, Lan, Danfeng
• Format: Journal Article
• Language: English
• Published: United States Hindawi 07.03.2023; John Wiley & Sons, Inc; Hindawi Limited
• Subjects: Analysis; Apoptosis; Cell growth; Cell proliferation; Chemokines; Cold; Crypts; CXCL10 protein; Cytokines; Disease; Epidermal growth factor; Epithelial cells; Epithelium; Ethylenediaminetetraacetic acid; Experiments; Gastrointestinal diseases; Goblet cells; Inflammation; Inflammatory bowel disease; Inflammatory bowel diseases; Inflammatory response; Interleukins; Intestine; Leucine; Monoclonal antibodies; Noggin protein; Organoids; Paneth cells; Pathogenesis; Proteins; Small intestine; Statistical significance; Stem cells; Structure-function relationships; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Canada; China; United Kingdom > UK; United States > US
• ISSN: 1687-966X; 1687-9678; 1687-9678
• DOI: 10.1155/2023/3328655

Intestinal epithelial dysfunction is critical in the development of inflammatory bowel disease (IBD). However, most cellular experiments related to epithelial barrier studies in IBD have been based on tumor cell line that lack a variety of intestinal epithelial cell types. Thus, intestinal organoids can present the three-dimensional structure and better simulate the physiological structure and function of the intestinal epithelium in vitro. Here, the crypts were isolated from the small intestine of mice; with the participation of major cytokines (EGF, Noggin, and R-Spondin 1 included), the intestinal organoids were established at a density of 100 crypts per well, containing intestinal stem cells (ISC), Paneth cells, goblet cells, and intestinal endocrine cells. We found that tumor necrosis factor-alpha (TNF-α) could induce the inflammatory response of intestinal organoids, and a dose of 10 ng/mL could maintain stable passaging of organoids for dynamic observation. After stimulation with TNF-α, the intestinal organoid cultures showed lower expression of the cell proliferation-related protein identified by monoclonal antibody Ki 67 (Ki67), the ISC marker leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5), and the intestinal tight junction proteins occludin (Ocln) and claudin-1 (Cldn1) while higher expression of the inflammatory cytokine interleukin- (IL-) 15 and the chemokines C-X-C motif ligand 2 (Cxcl2) and Cxcl10 significantly. In this study, we successfully established an epithelial inflammatory injury model of intestinal organoids, which provides an effective in vitro model for studying the pathogenesis and treatment of IBD.