title: Directed differentiation of cholangiocytes from human pluripotent stem cells.
abstract: Although bile duct disorders are well-recognized causes of liver disease, the molecular and cellular events leading to biliary dysfunction are poorly understood. To enable modeling and drug discovery for biliary disease, we describe a protocol that achieves efficient differentiation of biliary epithelial cells (cholangiocytes) from human pluripotent stem cells (hPSCs) through delivery of developmentally relevant cues, including NOTCH signaling. Using three-dimensional culture, the protocol yields cystic and/or ductal structures that express mature biliary markers, including apical sodium-dependent bile acid transporter, secretin receptor, cilia and cystic fibrosis transmembrane conductance regulator (CFTR). We demonstrate that hPSC-derived cholangiocytes possess epithelial functions, including rhodamine efflux and CFTR-mediated fluid secretion. Furthermore, we show that functionally impaired hPSC-derived cholangiocytes from cystic fibrosis patients are rescued by CFTR correctors. These findings demonstrate that mature cholangiocytes can be differentiated from hPSCs and used for studies of biliary development and disease.
authors.affiliation: 1] McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada. [2] Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada.
authors.forename: Mina
authors.initials: M
authors.lastname: Ogawa
authors.affiliation: McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada.
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ogawa
authors.affiliation: Program in Molecular Structure &Function, The Hospital for Sick Children Research Institute, Toronto, Ontario, Canada.
authors.forename: Christine E
authors.initials: CE
authors.lastname: Bear
authors.affiliation: Program in Molecular Structure &Function, The Hospital for Sick Children Research Institute, Toronto, Ontario, Canada.
authors.forename: Saumel
authors.initials: S
authors.lastname: Ahmadi
authors.affiliation: Program in Molecular Structure &Function, The Hospital for Sick Children Research Institute, Toronto, Ontario, Canada.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Chin
authors.affiliation: Department of Pediatrics, Oregon Health and Science University, Portland, Oregon, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Pediatrics, Oregon Health and Science University, Portland, Oregon, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Grompe
authors.affiliation: 1] McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada. [2] Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada. [3] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Gordon
authors.initials: G
authors.lastname: Keller
authors.affiliation: 1] Division of Gastroenterology, Hepatology and Nutrition, The Hospital for Sick Children, Toronto, Ontario, Canada. [2] Department of Pediatrics, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Binita M
authors.initials: BM
authors.lastname: Kamath
authors.affiliation: 1] Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada. [2] Division of General Surgery, University Health Network, Toronto, Ontario, Canada. [3] Department of Surgery, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Anand
authors.initials: A
authors.lastname: Ghanekar
mesh_terms: D001652:Bile Ducts / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology; D051880:Receptors, Notch / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D057905:Stem Cell Research
chemical_list: D051880:Receptors, Notch