Liver progenitor cells develop cholangiocyte-type epithelial polarity in three-dimensional culture.
Cholangiocytes are cellular components of the bile duct system of the liver, which originate from hepatoblasts during embryonic liver development. Although several transcription factors and signaling molecules have been implicated in bile duct development, its molecular mechanism has not been studied in detail. Here, we applied a three-dimensional (3D) culture technique to a liver progenitor cell line, HPPL, to establish an in vitro culture system in which HPPL acquire differentiated cholangiocyte characteristics. When HPPL were grown in a gel containing Matrigel, which contains extracellular matrix components of basement membrane, HPPL developed apicobasal polarity and formed cysts, which had luminal space inside. In the cysts, F-actin bundles and atypical protein kinase C were at the apical membrane, E-cadherin was localized at the lateral membrane, and beta-catenin and integrin alpha6 were located at the basolateral membrane. HPPL in cysts expressed cholangiocyte markers, including cytokeratin 19, integrin beta4, and aquaporin-1, but not a hepatocyte marker, albumin. Furthermore, HPPL transported rhodamine 123, a substrate for multidrug resistance gene products, from the basal side to the central lumen. These data indicate that HPPL develop cholangiocyte-type epithelial polarity in 3D culture. Phosphatidylinositol 3-kinase signaling was essential for proliferation and survival of HPPL in culture, whereas laminin-1 was a crucial component of Matrigel for inducing epithelial polarization of HPPL. Because HPPL cysts display structural and functional similarities with bile ducts, the 3D culture of HPPL recapitulates in vivo cholangiocyte differentiation and is useful to study the molecular mechanism of bile duct development in vitro.
Tanimizu
Naoki
N
Department of Anatomy, University of California San Francisco, San Francisco, CA 94143-2140, USA.
Miyajima
Atsushi
A
Mostov
Keith E
KE
D000818:Animals; D000831:Animals, Newborn; D001652:Bile Ducts / Q000166:cytology* / Q000196:embryology; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D018929:Cell Culture Techniques; D002460:Cell Line; D016764:Cell Polarity; D024042:Collagen Type I / Q000737:chemistry / Q000378:metabolism; D003560:Cysts / Q000473:pathology; D016207:Cytokines / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D053539:Keratin-19 / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000196:embryology / Q000254:growth & development; D008107:Liver Diseases / Q000473:pathology; D051379:Mice; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D015415:Biomarkers; D024042:Collagen Type I; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D053539:Keratin-19; D007797:Laminin; C121346:laminin 1