{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T01:43:13Z","timestamp":1784857393324,"version":"3.55.0"},"reference-count":23,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2024,11,26]],"date-time":"2024-11-26T00:00:00Z","timestamp":1732579200000},"content-version":"vor","delay-in-days":25,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2024,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Since their discovery, 3D cell cultures have emerged as powerful tools across various basic, translational research, and industrial discovery projects. One such application is in the physiological and pathophysiological modeling of pancreatic exocrine functions, which addresses critical clinical challenges, including acute and chronic pancreatitis. While several methods now exist for generating epithelial organoids (derived from induced pluripotent, embryonic, or adult stem cells), the advent of patient\u2010derived organoids (PDOs) with controlled polarity has introduced a new frontier in pancreatic research. This advancement has significantly expanded the methodological arsenal available for studying human pancreatic epithelial secretion.<\/jats:p><jats:p>In this article, we present basic protocols and a troubleshooting guide for an advanced culture method that results in an apical\u2010to\u2010basal polarity switch. Alongside the protocols, we emphasize a comprehensive cost breakdown, an aspect often challenging to estimate when implementing new techniques. By sharing the technical nuances and financial implications of these protocols, we aim to encourage researchers to transition from rodent models to primary human epithelial cells wherever feasible. This aligns with the U.S. Environmental Protection Agency's efforts to accelerate the translation of significant scientific findings to address major clinical needs. \u00a9 2024 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Establishment and maintenance of pancreatic PDOs<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Cryopreservation and thawing of pancreatic PDOs<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Inducing polarity switching in pancreatic PDOs<\/jats:p>","DOI":"10.1002\/cpz1.70045","type":"journal-article","created":{"date-parts":[[2024,11,26]],"date-time":"2024-11-26T16:01:33Z","timestamp":1732636893000},"source":"Crossref","is-referenced-by-count":3,"title":["Human Pancreas\u2010Derived Organoids with Controlled Polarity: Detailed Protocols and Experimental Timeline"],"prefix":"10.1002","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6404-1555","authenticated-orcid":false,"given":"Aletta","family":"Kiss","sequence":"first","affiliation":[{"name":"Department of Internal Medicine University of Szeged  Szeged Hungary"},{"name":"ELRN\u2010USZ Momentum Epithelial Cell Signaling and Secretion Research Group University of Szeged  Szeged Hungary"},{"name":"HCEMM\u2010USZ Molecular Gastroenterology Research Group University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Attila","family":"Farkas","sequence":"additional","affiliation":[{"name":"HCEMM\u2010USZ Functional Cell Biology and Immunology Advanced Core Facility University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ferhan","family":"Ayaydin","sequence":"additional","affiliation":[{"name":"HCEMM\u2010USZ Functional Cell Biology and Immunology Advanced Core Facility University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gy\u00f6rgy","family":"L\u00e1z\u00e1r","sequence":"additional","affiliation":[{"name":"Department of Surgery University of Szeged  Szeged Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2379-139X","authenticated-orcid":false,"given":"\u00c1rp\u00e1d","family":"Varga","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine University of Szeged  Szeged Hungary"},{"name":"ELRN\u2010USZ Momentum Epithelial Cell Signaling and Secretion Research Group University of Szeged  Szeged Hungary"},{"name":"HCEMM\u2010USZ Molecular Gastroenterology Research Group University of Szeged  Szeged Hungary"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5768-3090","authenticated-orcid":false,"given":"J\u00f3zsef","family":"Mal\u00e9th","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine University of Szeged  Szeged Hungary"},{"name":"ELRN\u2010USZ Momentum Epithelial Cell Signaling and Secretion Research Group University of Szeged  Szeged Hungary"},{"name":"HCEMM\u2010USZ Molecular Gastroenterology Research Group University of Szeged  Szeged Hungary"},{"name":"These authors contributed equally to this 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CRL\u20103276 (L\u2010WRN Cell Line)."},{"key":"e_1_2_9_3_3_1","unstructured":"https:\/\/www.atcc.org\/products\/crl\u20103276"},{"key":"e_1_2_9_3_4_1","unstructured":"The L\u2010WRN cell line is a crucial source of Wnt3a R\u2010spondin 3 and Noggin which are essential growth factors used in the culture of various organoid models including pancreatic organoids. This cell line provides conditioned medium that significantly enhances organoid growth maintenance and differentiation by supporting the stem cell niche. Understanding the use of the L\u2010WRN cell line is fundamental for replicating organoid culture methods as it underpins the conditions necessary for robust and reproducible organoid development."}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70045","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,2]],"date-time":"2024-12-02T04:57:52Z","timestamp":1733115472000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70045"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11]]},"references-count":23,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2024,11]]}},"alternative-id":["10.1002\/cpz1.70045"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70045","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11]]},"article-number":"e70045"}}