title: In vitro differentiation of hepatocyte-like cells from embryonic stem cells promoted by gene transfer of hepatocyte nuclear factor 3 beta.
abstract: We investigated the ability of a genetically altered embryonic stem (ES) cell line to promote endodermal differentiation toward hepatocytes in vitro by transfecting the hepatocyte nuclear factor-3beta (HNF-3beta) gene. Parental and HNF-3beta-transfected ES cells were initiated toward differentiation in embryoid bodies (EBs) for 5 days and the resulting EBs were transferred to an attached culture system. Albumin production was observed using an immuno-cytochemical method 7 days after induction of differentiation in almost all differentiating HNF-3beta-transfected ES cells, whereas scant immuno-reactivity against albumin was found on the same day in the cultures of differentiating parental ES cells. An analysis using a reverse transcriptase polymerase chain reaction revealed the HNF-4alpha expression in the HNF-3beta-transfected ES cells and also demonstrated that the expression of endodermal and hepatocyte-related markers, such as transthyretin, alpha-fetoprotein, albumin, alpha-1 antitrypsin, tryptophan-2,3-dioxygenase and phosphoenol-pyruvate carboxykinase, could be observed at an early stage in the outgrowths of HNF-3beta-transfected ES cells compared to the parental ES cells. These results suggest that HNF-3beta-transfected ES cells may be useful for the efficient induction of hepatocytes in vivo.
authors.affiliation: Department of Parasitology, Division of Developmental Biology, Nara Medical University, 840 Shijo-cho, Kashihara, 634-8521, Nara, Japan
authors.forename: Seiji
authors.initials: S
authors.lastname: Kanda
authors.forename: Akira
authors.initials: A
authors.lastname: Shiroi
authors.forename: Yukiteru
authors.initials: Y
authors.lastname: Ouji
authors.forename: Junichi
authors.initials: J
authors.lastname: Birumachi
authors.forename: Shigehiko
authors.initials: S
authors.lastname: Ueda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Fukui
authors.forename: Kouko
authors.initials: K
authors.lastname: Tatsumi
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Ishizaka
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Masahide
authors.initials: M
authors.lastname: Yoshikawa