title: Epithelial-mesenchymal transition delayed by E-cad to promote tissue formation in hepatic differentiation of mouse embryonic stem cells in vitro.
abstract: Hepatic differentiation of embryonic stem cells (ESCs) usually results in a single cell lineage, and the formation of liver tissues remains difficult. Here, we examine the role of epithelial-mesenchymal transition (EMT) that is regulated by epithelial cadherin (E-cad) expression in hepatic tissue formation from ESCs. E-cad was transfected into mouse ESCs to enable a stable expression of E-cad. Hepatic differentiation of ESCs was then induced by hepatic growth factors. Wnt/β-catenin signaling and EMT speed were examined to determine the differentiation process. Hepatic and angiogenesis markers, as well as differentiated cell-adhesive force were also examined to identify the hepatic tissue differentiation. In our results, E-cad expression gradually decreased in normal ESC (N-ESC) differentiation, but remained stable in the E-cad transfected ESC (EC-ESC) group. In EC-ESC differentiation, expressions of cytoplastic β-catenin and EMT were much lower and significantly prolonged. Angiogenesis markers vascular endothelial growth factor receptor-1 (VEGFR-1) and CD31/PECAM-1 were expressed only on day 5-13 in N-ESC differentiation, whereas VEGFR-1 and CD31/PECAM-1 were expressed prolonged on day 5-17 in the EC-ESC group and were coincident with the expression of hepatic markers. Finally, EC-ESC differentiation maintained multilayer-growth patterns, and abundant vascular network structures appeared and migrated in albumin-positive cell areas. The cellular adhesion forces between embryonic body cells in EC-ESC differentiation during day 13-17 were similar to those of mouse liver tissue. In conclusion, accelerated EMT due to the decreased E-cad expression may partially contribute to the failure of hepatic tissue formation in N-ESC differentiation. E-cad can act in synergy with hepatic growth factors and facilitate the early-stage formation of hepatic tissues through down-regulating Wnt/β-catenin signaling and delaying EMT. This work provides a new insight into hepatic tissue differentiation that is mediated by E-cad from ESC.
authors.affiliation: 1 Department of General Surgery, The First Affiliated Hospital, Sun Yat-sen University , Guangzhou, China .
authors.forename: Anbin
authors.initials: A
authors.lastname: Hu
authors.forename: Changzhen
authors.initials: C
authors.lastname: Shang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.forename: Nianfeng
authors.initials: N
authors.lastname: Sun
authors.forename: Linwei
authors.initials: L
authors.lastname: Wu
authors.forename: Yi
authors.initials: Y
authors.lastname: Ma
authors.forename: Xingyuan
authors.initials: X
authors.lastname: Jiao
authors.forename: Jun
authors.initials: J
authors.lastname: Min
authors.forename: Gucheng
authors.initials: G
authors.lastname: Zeng
authors.forename: Xiaoshun
authors.initials: X
authors.lastname: He
mesh_terms: D000818:Animals; D064200:Cdh1 Proteins / Q000378:metabolism*; D002448:Cell Adhesion; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000502:physiology*; D058750:Epithelial-Mesenchymal Transition; D015870:Gene Expression; D022781:Hepatocytes / Q000378:metabolism*; D008099:Liver / Q000166:cytology; D057507:Mice, 129 Strain; D008807:Mice, Inbred BALB C; D018919:Neovascularization, Physiologic / Q000235:genetics; D060449:Wnt Signaling Pathway
chemical_list: D064200:Cdh1 Proteins; C577585:Fzr1 protein, mouse