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However, the current fabrication techniques able to mimic the villus-crypt axis poses significant challenges in terms of reconstruction of the complex 3D microarchitecture. These challenges extend beyond mere structural intricacies to encompass the incorporation of diverse cell types and the management of intricate fluid dynamics within the system. Here, we introduce a novel microfluidic device called <jats:italic>In-Crypts<\/jats:italic>, which integrates a cell-instructive membrane aimed at inducing and guiding Caco-2 cells morphogenesis. Patterned topographical cues embossed onto the porous membrane induce the formation of a well-organized intestinal epithelium, characterized by proliferating crypt-like domains and differentiated villus-like regions. Notably, our cell-instructive porous membrane effectively sustains stem cells development, faithfully replicating the niche environment of <jats:italic>in vivo<\/jats:italic> intestinal crypts thus mirroring the cell biogeography observed <jats:italic>in vivo<\/jats:italic>. Moreover, by introducing dynamic fluid flow, we provide a faithful recapitulation of the native microenvironmental shear stress experienced by the intestinal epithelium. This stress plays a crucial role in influencing cell behaviour, differentiation, and overall functionality, thus offering a highly realistic model for studying intestinal physiology and pathology. The resulting intestinal epithelium exhibits significantly denser regions of mucus and microvilli, characteristic typically absent in static cultures, upregulating more than 1.5 of the amount expressed in the classical flattened configuration, enhanced epithelial cell differentiation and increased adsorptive surface area. Hence, the innovative design of <jats:italic>In-Crypts<\/jats:italic> proves the critical role of employing a cell-instructive membrane in argument the physiological relevance of organs-on-chips. This aspect, among others, will contribute to a more comprehensive understanding of organism function, directly impacting drug discovery and development.<\/jats:p>","DOI":"10.1088\/1758-5090\/ad6599","type":"journal-article","created":{"date-parts":[[2024,7,20]],"date-time":"2024-07-20T00:06:19Z","timestamp":1721433979000},"page":"045019","update-policy":"https:\/\/doi.org\/10.1088\/crossmark-policy","source":"Crossref","is-referenced-by-count":14,"title":["A novel membrane-on-chip guides morphogenesis for the reconstruction of the intestinal crypt-villus axis"],"prefix":"10.1088","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6469-1192","authenticated-orcid":true,"given":"Sara","family":"Sibilio","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raffaele","family":"Mennella","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7111-2961","authenticated-orcid":true,"given":"Vincenza De","family":"Gregorio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessia La","family":"Rocca","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Urciuolo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3500-0380","authenticated-orcid":true,"given":"Giorgia","family":"Imparato","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paolo A","family":"Netti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"266","published-online":{"date-parts":[[2024,8,12]]},"reference":[{"key":"bfad6599bib1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.261130","article-title":"\u2018Chip\u2019-ing away at morphogenesis\u2014application of organ-on-chip technologies to study tissue morphogenesis","volume":"136","author":"White","year":"2023","journal-title":"J. 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