title: Efficient hepatic differentiation of human induced pluripotent stem cells in a three-dimensional microscale culture.
abstract: Human induced pluripotent stem cells (iPSCs) represent a novel source of hepatocytes for drug development, disease modeling studies, and regenerative therapy for the treatment of liver diseases. A number of protocols for generating functional hepatocytes have been reported worldwide; however, reproducible and efficient differentiation remained challenging under conventional two-dimensional (2D) culture. In this study, we describe an efficient differentiation protocol for generating functional hepatocyte-like cells from human iPSC-derived homogenous hepatic endoderm cells combined with three-dimensional (3D) microscale culture system. First, hepatic endoderm cells (iPSC-HEs) were directly differentiated using two-step approaches, and then cultured in the 3D micropattern plate. Human iPSC-HEs quickly reaggregated and formed hundreds of round-shaped spheroids at day 4 of cell plating. The size distribution of iPSC-HEs derived spheroids was relatively uniform around 100-200 μm in diameter. After 14 days, iPSC-HEs efficiently differentiated into hepatocyte-like cells in terms of hepatic maker gene expression compared with conventional 2D approach. We conclude that our scalable and three-dimensional culture system would be one promising approach to generate a huge number of hepatocyte-like cells from human iPSCs aiming at future industrial and clinical applications.
authors.affiliation: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, 3-9, Fukuura, Kanazawa-ku, Yokohama, Kanagawa, 236-0004, Japan.
authors.forename: Ran-Ran
authors.initials: RR
authors.lastname: Zhang
authors.forename: Takanori
authors.initials: T
authors.lastname: Takebe
authors.forename: Leina
authors.initials: L
authors.lastname: Miyazaki
authors.forename: Maho
authors.initials: M
authors.lastname: Takayama
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Koike
authors.forename: Masaki
authors.initials: M
authors.lastname: Kimura
authors.forename: Masahiro
authors.initials: M
authors.lastname: Enomura
authors.forename: Yun-Wen
authors.initials: YW
authors.lastname: Zheng
authors.forename: Keisuke
authors.initials: K
authors.lastname: Sekine
authors.forename: Hideki
authors.initials: H
authors.lastname: Taniguchi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice