title: Efficient generation of hepatoblasts from human ES cells and iPS cells by transient overexpression of homeobox gene HEX.
abstract: Human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have the potential to differentiate into all cell lineages, including hepatocytes, in vitro. Induced hepatocytes have a wide range of potential application in biomedical research, drug discovery, and the treatment of liver disease. However, the existing protocols for hepatic differentiation of PSCs are not very efficient. In this study, we developed an efficient method to induce hepatoblasts, which are progenitors of hepatocytes, from human ESCs and iPSCs by overexpression of the HEX gene, which is a homeotic gene and also essential for hepatic differentiation, using a HEX-expressing adenovirus (Ad) vector under serum/feeder cell-free chemically defined conditions. Ad-HEX-transduced cells expressed α-fetoprotein (AFP) at day 9 and then expressed albumin (ALB) at day 12. Furthermore, the Ad-HEX-transduced cells derived from human iPSCs also produced several cytochrome P450 (CYP) isozymes, and these P450 isozymes were capable of converting the substrates to metabolites and responding to the chemical stimulation. Our differentiation protocol using Ad vector-mediated transient HEX transduction under chemically defined conditions efficiently generates hepatoblasts from human ESCs and iPSCs. Thus, our methods would be useful for not only drug screening but also therapeutic applications.
authors.affiliation: Department of Biochemistry and Molecular Biology, Osaka University, Osaka, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Inamura
authors.forename: Kenji
authors.initials: K
authors.lastname: Kawabata
authors.forename: Kazuo
authors.initials: K
authors.lastname: Takayama
authors.forename: Katsuhisa
authors.initials: K
authors.lastname: Tashiro
authors.forename: Fuminori
authors.initials: F
authors.lastname: Sakurai
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Katayama
authors.forename: Masashi
authors.initials: M
authors.lastname: Toyoda
authors.forename: Hidenori
authors.initials: H
authors.lastname: Akutsu
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Miyagawa
authors.forename: Hajime
authors.initials: H
authors.lastname: Okita
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Kiyokawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
authors.forename: Takao
authors.initials: T
authors.lastname: Hayakawa
authors.forename: Miho K
authors.initials: MK
authors.lastname: Furue
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mizuguchi
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005801:Genes, Homeobox / Q000235:genetics / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*
chemical_list: C078492:HHEX protein, human; D018398:Homeodomain Proteins; D014157:Transcription Factors; D003577:Cytochrome P-450 Enzyme System