title: Liver Regeneration Using Cultured Liver Bud.
abstract: Here, we describe a protocol to develop a three-dimensional (3D) liver bud-like tissue from human iPSCs in vitro. This method mainly consists of two parts: (1) hepatic endoderm (HE) differentiation from human iPSCs in 2D culture and (2) co-culturing iPSC-HE with endothelial and mesenchymal cells. First, iPSCs were differentiated into definitive endoderm (DE) cells, and the DE cells were differentiated into HE cells, which were then co-cultured with endothelial cells and mesenchymal cells on Matrigel-coated plastic plates or micropattern plates. The cells rapidly condensed to generate 3D tissue masses. We named these iPSC liver buds (iPSC-LBs) because they resemble the developing liver bud from the perspective of gene expression, cell proliferation, and cell proportion. This liver bud culture system provides a novel approach for future clinical applications, for drug development, and as a tool for studying human development.
authors.affiliation: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku 3-9, Yokohama, Kanagawa, 236-0004, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Sekine
authors.affiliation: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku 3-9, Yokohama, Kanagawa, 236-0004, Japan.|Advanced medical research center, Yokohama City University, Kanazawa-ku 3-9, Yokohama, Kanagawa, 236-0004, Japan.|PRESTO, Japan Science and Technology Agency, 4-1-8, Honcho, Kawaguchi, Saitama, 332-0012, Japan.|Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, 3333 Burnet Avenue, Cincinnati, OH, 45229-3039, USA.
authors.forename: Takanori
authors.initials: T
authors.lastname: Takebe
authors.affiliation: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku 3-9, Yokohama, Kanagawa, 236-0004, Japan. rtanigu@yokohama-cu.ac.jp.|Advanced medical research center, Yokohama City University, Kanazawa-ku 3-9, Yokohama, Kanagawa, 236-0004, Japan. rtanigu@yokohama-cu.ac.jp.
authors.forename: Hideki
authors.initials: H
authors.lastname: Taniguchi
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D004707:Endoderm / Q000166:cytology; D042783:Endothelial Cells / Q000166:cytology; D022781:Hepatocytes / Q000166:cytology; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D008099:Liver / Q000166:cytology*; D008115:Liver Regeneration / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods
keywords: Drug development; Human-induced pluripotent stem cells; Liver bud; Multicellular interaction; Regenerative medicine; Self-organization; Three-dimensional culture