title: Paracrine signals regulate human liver organoid maturation from induced pluripotent stem cells.
abstract: A self-organizing organoid model provides a new approach to study the mechanism of human liver organogenesis. Previous animal models documented that simultaneous paracrine signaling and cell-to-cell surface contact regulate hepatocyte differentiation. To dissect the relative contributions of the paracrine effects, we first established a liver organoid using human induced pluripotent stem cells (iPSCs), mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs) as previously reported. Time-lapse imaging showed that hepatic-specified endoderm iPSCs (HE-iPSCs) self-assembled into three-dimensional organoids, resulting in hepatic gene induction. Progressive differentiation was demonstrated by hepatic protein production afterin vivoorganoid transplantation. To assess the paracrine contributions, we employed a Transwell system in which HE-iPSCs were separately co-cultured with MSCs and/or HUVECs. Although the three-dimensional structure did not form, their soluble factors induced a hepatocyte-like phenotype in HE-iPSCs, resulting in the expression of bile salt export pump. In conclusion, the mesoderm-derived paracrine signals promote hepatocyte maturation in liver organoids, but organoid self-organization requires cell-to-cell surface contact. Ourin vitromodel demonstrates a novel approach to identify developmental paracrine signals regulating the differentiation of human hepatocytes.
authors.affiliation: Pediatric Liver Care Center, Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA akihiro.asai@cchmc.org.
authors.forename: Akihiro
authors.identifier: 0000-0003-3029-5754
authors.initials: A
authors.lastname: Asai
authors.affiliation: Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, OH 45229, USA.
authors.forename: Eitaro
authors.initials: E
authors.lastname: Aihara
authors.affiliation: Division of Pediatric Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Carey
authors.initials: C
authors.lastname: Watson
authors.affiliation: Pediatric Liver Care Center, Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Reena
authors.initials: R
authors.lastname: Mourya
authors.affiliation: Pediatric Liver Care Center, Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Tatsuki
authors.initials: T
authors.lastname: Mizuochi
authors.affiliation: Pediatric Liver Care Center, Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Pranavkumar
authors.initials: P
authors.lastname: Shivakumar
authors.affiliation: Pediatric Liver Care Center, Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Kieran
authors.initials: K
authors.lastname: Phelan
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Mayhew
authors.affiliation: Division of Pediatric Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Helmrath
authors.affiliation: Department of Regenerative Medicine, Yokohama City University, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Takebe
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: James
authors.initials: J
authors.lastname: Wells
authors.affiliation: Pediatric Liver Care Center, Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Bezerra
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism; D001692:Biological Transport; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D016764:Cell Polarity; D018920:Coculture Techniques; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000166:cytology / Q000648:ultrastructure; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D009024:Morphogenesis / Q000235:genetics; D009928:Organ Specificity / Q000235:genetics; D009940:Organoids / Q000166:cytology* / Q000378:metabolism; D019899:Paracrine Communication; D011506:Proteins / Q000032:analysis
chemical_list: D001647:Bile Acids and Salts; D015415:Biomarkers; D011506:Proteins
keywords: ABCB11; Hepatocyte differentiation; Liver development; Regeneration; Tissue engineering