{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T09:11:17Z","timestamp":1784365877948,"version":"3.55.0"},"reference-count":46,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T00:00:00Z","timestamp":1768867200000},"content-version":"vor","delay-in-days":19,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01DK119435"],"award-info":[{"award-number":["R01DK119435"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01DK103775"],"award-info":[{"award-number":["R01DK103775"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Physiological Reports"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Cultures of primary mouse bile duct epithelial cells are a valuable tool to study cholangiocyte secretion and bile formation. However, freshly isolated cells have a limited ability to expand in culture. Here we report a novel isolation and culture technique for normal mouse cholangiocytes (NMC) that enables long\u2010term growth without compromising function. Mouse cholangiocytes were isolated and cultured in conditioned medium (CM) that was subsequently supplemented with ROCK inhibitor Y\u201027632. Expression of cholangiocyte markers was assessed by qPCR, immunofluorescence, and Western blotting. Patch clamp techniques were used to measure cAMP\u2010activated Cl\u2010 current, Ca2+\u2212activated Cl\u2010 current, and volume\u2010stimulated Cl\u2010 current. We obtained NMC cultures that were polarized and maintained a cholangiocyte phenotype for over 50 passages. Functional studies show that ion channel activity is maintained in NMC regardless of the number of passages and despite removal of CM. NMC also perform other physiological functions such as ATP release and intracellular Ca2+ changes in response to stimulation with bile acids. Thus, our isolation procedure produces viable NMC that maintain biophysical properties in long\u2010term culture. We also demonstrate the utility of NMC in studies investigating the cellular mechanisms responsible for cholangiocyte secretion and bile formation.<\/jats:p>","DOI":"10.14814\/phy2.70732","type":"journal-article","created":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T05:31:05Z","timestamp":1768973465000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation and long\u2010term culture of primary mouse cholangiocytes that retain biophysical properties and distinct Cl\u2010conductances: An initial study"],"prefix":"10.14814","volume":"14","author":[{"given":"Qin","family":"Li","sequence":"first","affiliation":[{"name":"Department of Pediatrics University of Pittsburgh Medical Center (UPMC)  Pittsburgh Pennsylvania USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youxue","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University of Texas Southwestern Medical Center  Dallas Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristy","family":"Boggs","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University of Pittsburgh Medical Center (UPMC)  Pittsburgh Pennsylvania USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles","family":"Kresge","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University of Texas Southwestern Medical Center  Dallas Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6051-5338","authenticated-orcid":false,"given":"Kari","family":"Nejak\u2010Bowen","sequence":"additional","affiliation":[{"name":"Organ Pathobiology and Therapeutics Institute, University of Pittsburgh School of Medicine  Pittsburgh Pennsylvania USA"},{"name":"Department of Pharmacology and Chemical Biology University of Pittsburgh School of Medicine, Pittsburgh  Pittsburgh Pennsylvania USA"},{"name":"Pittsburgh Liver Research Center, University of Pittsburgh and University of Pittsburgh Medical Center  Pittsburgh Pennsylvania USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,1,20]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41575-019-0125-y"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.2040-4603.2013.tb00519.x"},{"key":"e_1_2_12_4_1","doi-asserted-by":"crossref","first-page":"2619","DOI":"10.1172\/JCI42297","article-title":"Human keratinocytes are efficiently immortalized by a rho kinase inhibitor","volume":"120","author":"Chapman S.","year":"2010","journal-title":"The Journal of Clinical Investigation"},{"key":"e_1_2_12_5_1","doi-asserted-by":"crossref","DOI":"10.3389\/fmicb.2022.907901","article-title":"Highly efficient methods to culture mouse Cholangiocytes and small intestine organoids","volume":"13","author":"Chen W.","year":"2022","journal-title":"Frontiers in Microbiology"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1155\/2019\/3010656"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.006064"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.164970"},{"key":"e_1_2_12_9_1","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1172\/JCI200112190","article-title":"p38 MAP kinase modulates liver cell volume through inhibition of membrane Na+ permeability","volume":"108","author":"Feranchak A. 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