Directed differentiation of cholangiocytes from human pluripotent stem cells

• Published in: Nature biotechnology Vol. 33; no. 8; pp. 853 - 861
• Main Authors: Ogawa, Mina, Ogawa, Shinichiro, Bear, Christine E, Ahmadi, Saumel, Chin, Stephanie, Li, Bin, Grompe, Markus, Keller, Gordon, Kamath, Binita M, Ghanekar, Anand
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.08.2015; Nature Publishing Group
• Subjects: 13; 13/100; 13/106; 631/136/1425; 631/532/2064/2158; 631/61/2320; 692/698/2741/288; 692/699/1503/1328/1325; Agriculture; Bile ducts; Bile Ducts - cytology; Bioinformatics; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Cell Culture Techniques - methods; Cell differentiation; Cell Differentiation - physiology; Cells, Cultured; Common bile duct; Epithelial cells; Epithelial Cells - cytology; Epithelial Cells - physiology; Humans; Life Sciences; Liver diseases; Pharmacology; Physiological aspects; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - physiology; Receptors, Notch - metabolism; Signal Transduction - physiology; Sodium; Stem Cell Research; Stem cells
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt.3294

A protocol for generating biliary epithelial cells from human pluripotent stem cells facilitates disease modeling and drug screening. 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.