Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures
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...
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| Format: | Article |
| Language: | English |
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Wiley
2023-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2023/3328655 |
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| author | Chengfeng Xing Guili Liang Xin Yu Anxing Zhang Xiang Luo Yu Liu Zengli Tang Bian Wu Zhengji Song Danfeng Lan |
| author_facet | Chengfeng Xing Guili Liang Xin Yu Anxing Zhang Xiang Luo Yu Liu Zengli Tang Bian Wu Zhengji Song Danfeng Lan |
| author_sort | Chengfeng Xing |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-8413486f884b4f90b883e1bfa50b3b97 |
| institution | Kabale University |
| issn | 1687-9678 |
| language | English |
| publishDate | 2023-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-8413486f884b4f90b883e1bfa50b3b972025-08-20T03:37:46ZengWileyStem Cells International1687-96782023-01-01202310.1155/2023/3328655Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid CulturesChengfeng Xing0Guili Liang1Xin Yu2Anxing Zhang3Xiang Luo4Yu Liu5Zengli Tang6Bian Wu7Zhengji Song8Danfeng Lan9Department of GastroenterologyKunming Medical UniversityMedical SchoolDepartment of GastroenterologyState Key Laboratory of Primate Biomedical ResearchDepartment of GastroenterologyState Key Laboratory of Primate Biomedical ResearchDepartment of General Surgery IIDepartment of GastroenterologyDepartment of GastroenterologyIntestinal 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.http://dx.doi.org/10.1155/2023/3328655 |
| spellingShingle | Chengfeng Xing Guili Liang Xin Yu Anxing Zhang Xiang Luo Yu Liu Zengli Tang Bian Wu Zhengji Song Danfeng Lan Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures Stem Cells International |
| title | Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures |
| title_full | Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures |
| title_fullStr | Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures |
| title_full_unstemmed | Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures |
| title_short | Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures |
| title_sort | establishment of epithelial inflammatory injury model using intestinal organoid cultures |
| url | http://dx.doi.org/10.1155/2023/3328655 |
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