title: The promotion of hepatic maturation of human pluripotent stem cells in 3D co-culture using type I collagen and Swiss 3T3 cell sheets.
abstract: Hepatocyte-like cells differentiated from human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) are known to be a useful cell source for drug screening. We recently developed an efficient hepatic differentiation method from hESCs and hiPSCs by sequential transduction of FOXA2 and HNF1α. It is known that the combination of three-dimensional (3D) culture and co-culture, namely 3D co-culture, can maintain the functions of primary hepatocytes. However, hepatic maturation of hESC- or hiPSC-derived hepatocyte-like cells (hEHs or hiPHs, respectively) by 3D co-culture systems has not been examined. Therefore, we utilized a cell sheet engineering technology to promote hepatic maturation. The gene expression levels of hepatocyte-related markers (such as cytochrome P450 enzymes and conjugating enzymes) and the amount of albumin secretion in the hEHs or hiPHs, which were 3D co-cultured with the Swiss 3T3 cell sheet, were significantly up-regulated in comparison with those in the hEHs or hiPHs cultured in a monolayer. Furthermore, we found that type I collagen synthesized in Swiss 3T3 cells plays an important role in hepatic maturation. The hEHs or hiPHs that were 3D co-cultured with the Swiss 3T3 cell sheet would be powerful tools for medical applications, such as drug screening.
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Nagamoto
authors.forename: Katsuhisa
authors.initials: K
authors.lastname: Tashiro
authors.forename: Kazuo
authors.initials: K
authors.lastname: Takayama
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ohashi
authors.forename: Kenji
authors.initials: K
authors.lastname: Kawabata
authors.forename: Fuminori
authors.initials: F
authors.lastname: Sakurai
authors.forename: Masashi
authors.initials: M
authors.lastname: Tachibana
authors.forename: Takao
authors.initials: T
authors.lastname: Hayakawa
authors.forename: Miho Kusuda
authors.initials: MK
authors.lastname: Furue
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mizuguchi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods*; D024042:Collagen Type I / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D008099:Liver / Q000166:cytology*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology*; D041701:Swiss 3T3 Cells
chemical_list: D024042:Collagen Type I