Dotti, IsabellaMayorgas, AidaSalas Martínez, Azucena2023-07-262023-07-262022-10-271932-6203https://hdl.handle.net/2445/201181Ulcerative 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.13 p.application/pdfengcc by (c) Dotti, Isabella et al, 2022http://creativecommons.org/licenses/by/3.0/es/Malalties inflamatòries intestinalsMembrana mucosaInflammatory bowel diseasesMucous membraneinfo:eu-repo/semantics/article2023-07-10info:eu-repo/semantics/openAccess933150236301950