title: Expression of Tight Junction Components in Hepatocyte-Like Cells Differentiated from Human Embryonic Stem Cells.
abstract: Human embryonic stem cells can be differentiated in vitro into a wide variety of progeny cells by addition of different morphogens and growth factors. Our aim was to monitor the expression pattern of tight junction (TJ) components and various cellular markers during differentiation of stem cell lines toward the hepatic lineage. Human embryonic stem cell lines (HUES1, HUES9) were differentiated into endoderm-like cells, and further differentiated to hepatocyte-like cells. Gene expressions of Oct3/4, Nanog, alpha-fetoprotein, albumin, cytokeratins (CK-7, CK-8, CK-18, CK-19), ATP-binding cassette (ABC) transporters (ABCC2, ABCC7, ABCG2), and various TJ components, including claudin-1, claudin-4, claudin-5, claudin-7, and tricellulin, as well as an extracellular matrix component, agrin were monitored during hepatic differentiation by real-time quantitative PCR. The differentiated cells exhibit epithelial morphology and functional assessments similar to that of hepatocytes. The expression level of stem cell marker genes (Oct3/4 and Nanog) significantly and gradually decreased, while liver-associated genes (alpha-fetoprotein, albumin) reached their highest expression at the end of the differentiation. The endoderm-like cells expressed claudin-1, which declined eventually. The expression levels of cholangiocyte markers including claudin-4, CK-7, CK-19, and agrin gradually increased and reached their highest level at the final stage of differentiation. In contrast, these cells did not express notable level of claudin-7, CK-8 and tricellulin. The marker set used for monitoring differentiation revealed both hepatocyte and cholangiocyte characteristics of the differentiated cells at the final stage. This is the first report describing the expression level changes of various TJ components, and underlining their importance in hepatic differentiation.
authors.affiliation: 2nd Department of Pathology, Semmelweis University, Üllői Street 93, 1091, Budapest, Hungary.
authors.forename: Boglárka
authors.initials: B
authors.lastname: Erdélyi-Belle
authors.forename: György
authors.initials: G
authors.lastname: Török
authors.forename: Ágota
authors.initials: Á
authors.lastname: Apáti
authors.forename: Balázs
authors.initials: B
authors.lastname: Sarkadi
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Schaff
authors.forename: András
authors.initials: A
authors.lastname: Kiss
authors.forename: László
authors.initials: L
authors.lastname: Homolya
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D062445:Claudin-1 / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000378:metabolism*; D004707:Endoderm / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D000089762:Multidrug Resistance-Associated Protein 2; D019108:Tight Junctions / Q000235:genetics* / Q000378:metabolism*
chemical_list: C000715087:ABCC2 protein, human; D015415:Biomarkers; D062445:Claudin-1; D000089762:Multidrug Resistance-Associated Protein 2
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