title: Three-dimensional culture and cAMP signaling promote the maturation of human pluripotent stem cell-derived hepatocytes.
abstract: Human pluripotent stem cells (hPSCs) represent a novel source of hepatocytes for drug metabolism studies and cell-based therapy for the treatment of liver diseases. These applications are, however, dependent on the ability to generate mature metabolically functional cells from the hPSCs. Reproducible and efficient generation of such cells has been challenging to date, owing to the fact that the regulatory pathways that control hepatocyte maturation are poorly understood. Here, we show that the combination of three-dimensional cell aggregation and cAMP signaling enhance the maturation of hPSC-derived hepatoblasts to a hepatocyte-like population that displays expression profiles and metabolic enzyme levels comparable to those of primary human hepatocytes. Importantly, we also demonstrate that generation of the hepatoblast population capable of responding to cAMP is dependent on appropriate activin/nodal signaling in the definitive endoderm at early stages of differentiation. Together, these findings provide new insights into the pathways that regulate maturation of hPSC-derived hepatocytes and in doing so provide a simple and reproducible approach for generating metabolically functional cell populations.
authors.affiliation: McEwen Centre For Regenerative Medicine, University Health Network, Toronto, ON M5G 1L7, Canada.
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ogawa
authors.forename: James
authors.initials: J
authors.lastname: Surapisitchat
authors.forename: Carl
authors.initials: C
authors.lastname: Virtanen
authors.forename: Mina
authors.initials: M
authors.lastname: Ogawa
authors.forename: Maryam
authors.initials: M
authors.lastname: Niapour
authors.forename: Kim S
authors.initials: KS
authors.lastname: Sugamori
authors.forename: Shuang
authors.initials: S
authors.lastname: Wang
authors.forename: Laura
authors.initials: L
authors.lastname: Tamblyn
authors.forename: Chantal
authors.initials: C
authors.lastname: Guillemette
authors.forename: Ewa
authors.initials: E
authors.lastname: Hoffmann
authors.forename: Bin
authors.initials: B
authors.lastname: Zhao
authors.forename: Stephen
authors.initials: S
authors.lastname: Strom
authors.forename: Rebecca R
authors.initials: RR
authors.lastname: Laposa
authors.forename: Rachel F
authors.initials: RF
authors.lastname: Tyndale
authors.forename: Denis M
authors.initials: DM
authors.lastname: Grant
authors.forename: Gordon
authors.initials: G
authors.lastname: Keller
mesh_terms: D028341:Activins / Q000378:metabolism; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D000242:Cyclic AMP / Q000378:metabolism*; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D055457:Nodal Protein / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D015398:Signal Transduction; D059467:Transcriptome
chemical_list: C526007:NODAL protein, human; D055457:Nodal Protein; D028341:Activins; D000242:Cyclic AMP
keywords: Endoderm; Hepatocyte; Human pluripotent stem cell; Maturation; Three dimensional culture; cAMP signaling