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Although frequent <jats:italic>KRAS<\/jats:italic> mutations and epigenetic changes at the <jats:italic>INK4A\/ARF<\/jats:italic> locus have been identified, the molecular pathogenesis of BTC is unclear and the development of corresponding anticancer agents remains inadequate. We isolated epithelial cell adhesion molecule (EpCAM)\u2013positive BECs from the mouse intrahepatic bile duct, gallbladder, and extrahepatic bile duct, and established organoids derived from these cells. Introduction of activated KRAS and homozygous deletion of <jats:italic>Ink4a\/Arf<\/jats:italic> in the cells of each organoid type conferred the ability to form lethal metastatic adenocarcinoma with differentiated components and a pronounced desmoplastic reaction on cell transplantation into syngeneic mice, indicating that the manipulated cells correspond to BTC\u2013initiating cells. The syngeneic mouse models recapitulate the pathological features of human IHCC, GC, and EHCC, and they should therefore prove useful for the investigation of BTC carcinogenesis and the development of new therapeutic strategies. Tumor cells isolated from primary tumors formed organoids in three\u2010dimensional culture, and serial syngeneic transplantation of these cells revealed that their cancer stem cell properties were supported by organoid culture, but not by adherent culture. Adherent culture thus attenuated tumorigenic activity as well as the expression of both epithelial and stem cell markers, whereas the expression of epithelial\u2010mesenchymal transition (EMT)\u2013related transcription factor genes and mesenchymal cell markers was induced. Our data show that organoid culture is important for maintenance of epithelial cell characteristics, stemness, and tumorigenic activity of BTC\u2013initiating cells.<\/jats:p>","DOI":"10.1111\/cas.14703","type":"journal-article","created":{"date-parts":[[2020,10,17]],"date-time":"2020-10-17T11:53:32Z","timestamp":1602935612000},"page":"1822-1838","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Oncogenic KRAS\u2013expressing organoids with biliary epithelial stem cell properties give rise to biliary tract cancer in mice"],"prefix":"10.1111","volume":"112","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2234-1135","authenticated-orcid":false,"given":"Akiyoshi","family":"Kasuga","sequence":"first","affiliation":[{"name":"Division of Gene Regulation Institute for Advanced Medical Research Keio University School of Medicine Tokyo Japan"},{"name":"Division of Gastroenterology and Hepatology Department of Internal 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