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Title: | Generation of human colon organoids from healthy and inflammatory bowel disease mucosa |
Author: | Dotti, Isabella Mayorgas, Aida Salas, Azucena |
Keywords: | Malalties inflamatòries intestinals Membrana mucosa Inflammatory bowel diseases Mucous membrane |
Issue Date: | 27-Oct-2022 |
Abstract: | Ulcerative colitis and Crohn's disease are chronic inflammatory bowel diseases (IBD) of unknown cause characterized by a relapsing-remitting behavior. Growing evidence supports the idea that the epithelial barrier plays a central role in the pathogenesis of IBD as well as in its evolution over time, thus representing a potential target for novel therapeutic options. In the last decade, the introduction of 3D epithelial cultures from ex vivo-expanded intestinal adult stem cells (ASCs) has impacted our ability to study the function of the epithelium in several gastrointestinal disorders, including IBD. Here, we describe in detail a reproducible protocol to generate Matrigel-embedded epithelial organoids from ASCs of non-IBD and IBD donors using small colonic biopsies, including steps for its optimization. A slightly modified version of this protocol is also provided in case surgical samples are used. With this method, epithelial organoids can be expanded over several passages, thereby generating a large quantity of viable cells that can be used in multiple downstream analyses including genetic, transcriptional, proteomic and/or functional studies. In addition, 3D cultures generated using our protocol are suitable for the establishment of 2D cultures, which can model relevant cell-to-cell interactions that occur in IBD mucosa. |
Note: | Reproducció del document publicat a: https://doi.org/10.1371/journal.pone.0276195 |
It is part of: | Plos One, 2022, vol. 17, num. 10 |
URI: | http://hdl.handle.net/2445/201181 |
Related resource: | https://doi.org/10.1371/journal.pone.0276195 |
ISSN: | 1932-6203 |
Appears in Collections: | Articles publicats en revistes (IDIBAPS: Institut d'investigacions Biomèdiques August Pi i Sunyer) |
Files in This Item:
File | Description | Size | Format | |
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Generation of human colon organoids from healthy and inflammatory bowel disease mucosa_PlosOne.pdf | 2.7 MB | Adobe PDF | View/Open |
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