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Organoids can consist of all intestinal cell types and are supposed to better reflect the <jats:italic>in vivo<\/jats:italic> situation. Growing three\u2010dimensionally, with the apical side facing the lumen, application of typical physiological techniques to analyze the barrier is difficult. Organoid\u2010derived monolayers (ODMs) were developed to overcome this. After optimizing culturing conditions, ODMs were characterized and compared to Caco\u20102 and duodenal tissue. Tight junction composition and appearance were analyzed, and electrophysiological barrier properties, like paracellular and transcellular barrier function and macromolecule permeability, were evaluated. Furthermore, transcriptomic data were analyzed. ODMs had tight junction protein expression and paracellular barrier properties much more resembling the originating tissue than Caco\u20102. Transcellular barrier was similar between ODMs and native tissue but was increased in Caco\u20102. Transcriptomic data showed that Caco\u20102 expressed fewer solute carriers than ODMs and native tissue. In conclusion, while Caco\u20102 cells differ mostly in transcellular properties, ODMs reflect trans\u2010 and paracellular properties of the originating tissue. If cultured under optimized conditions, ODMs possess reproducible functionality, and the variety of different cell types makes them a suitable model for human tissue\u2010specific investigations.<\/jats:p>","DOI":"10.1111\/nyas.14804","type":"journal-article","created":{"date-parts":[[2022,6,6]],"date-time":"2022-06-06T19:40:37Z","timestamp":1654544437000},"page":"155-167","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Human duodenal organoid\u2010derived monolayers serve as a suitable barrier model for duodenal tissue"],"prefix":"10.1111","volume":"1515","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5069-9628","authenticated-orcid":false,"given":"Franziska","family":"Wei\u00df","sequence":"first","affiliation":[{"name":"Clinical Physiology\/Nutritional Medicine Charit\u00e9 \u2013 Universit\u00e4tsmedizin Berlin, CBF  Berlin 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