Generation of human colon organoids from healthy and inflammatory bowel disease mucosa.
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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2022-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0276195 |
| _version_ | 1797989980083060736 |
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| author | Isabella Dotti Aida Mayorgas Azucena Salas |
| author_facet | Isabella Dotti Aida Mayorgas Azucena Salas |
| author_sort | Isabella Dotti |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T08:29:17Z |
| format | Article |
| id | doaj.art-ae6f2b8a3e3a4585ab928257334eb8b9 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-11T08:29:17Z |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ae6f2b8a3e3a4585ab928257334eb8b92022-12-22T04:34:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-011710e027619510.1371/journal.pone.0276195Generation of human colon organoids from healthy and inflammatory bowel disease mucosa.Isabella DottiAida MayorgasAzucena SalasUlcerative 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.https://doi.org/10.1371/journal.pone.0276195 |
| spellingShingle | Isabella Dotti Aida Mayorgas Azucena Salas Generation of human colon organoids from healthy and inflammatory bowel disease mucosa. PLoS ONE |
| title | Generation of human colon organoids from healthy and inflammatory bowel disease mucosa. |
| title_full | Generation of human colon organoids from healthy and inflammatory bowel disease mucosa. |
| title_fullStr | Generation of human colon organoids from healthy and inflammatory bowel disease mucosa. |
| title_full_unstemmed | Generation of human colon organoids from healthy and inflammatory bowel disease mucosa. |
| title_short | Generation of human colon organoids from healthy and inflammatory bowel disease mucosa. |
| title_sort | generation of human colon organoids from healthy and inflammatory bowel disease mucosa |
| url | https://doi.org/10.1371/journal.pone.0276195 |
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