title: Differentiation and transplantation of human embryonic stem cell-derived hepatocytes.
abstract: OBJECTIVE
  The ability to obtain unlimited numbers of human hepatocytes would improve the development of cell-based therapies for liver diseases, facilitate the study of liver biology, and improve the early stages of drug discovery. Embryonic stem cells are pluripotent, potentially can differentiate into any cell type, and therefore could be developed as a source of human hepatocytes.
  
  
  METHODS
  To generate human hepatocytes, human embryonic stem cells were differentiated by sequential culture in fibroblast growth factor 2 and human activin-A, hepatocyte growth factor, and dexamethasone. Functional hepatocytes were isolated by sorting for surface asialoglycoprotein-receptor expression. Characterization was performed by real-time polymerase chain reaction, immunohistochemistry, immunoblot, functional assays, and transplantation.
  
  
  RESULTS
  Embryonic stem cell-derived hepatocytes expressed liver-specific genes, but not genes representing other lineages, secreted functional human liver-specific proteins similar to those of primary human hepatocytes, and showed human hepatocyte cytochrome P450 metabolic activity. Serum from rodents given injections of embryonic stem cell-derived hepatocytes contained significant amounts of human albumin and alpha1-antitrypsin. Colonies of cytokeratin-18 and human albumin-expressing cells were present in the livers of recipient animals.
  
  
  CONCLUSIONS
  Human embryonic stem cells can be differentiated into cells with many characteristics of primary human hepatocytes. Hepatocyte-like cells can be enriched and recovered based on asialoglycoprotein-receptor expression and potentially could be used in drug discovery research and developed as therapeutics.
authors.affiliation: Department of Surgery, University of Nebraska Medical Center, Omaha, Nebraska, USA.
authors.forename: Hesham
authors.initials: H
authors.lastname: Basma
authors.forename: Alejandro
authors.initials: A
authors.lastname: Soto-Gutiérrez
authors.forename: Govardhana Rao
authors.initials: GR
authors.lastname: Yannam
authors.forename: Liping
authors.initials: L
authors.lastname: Liu
authors.forename: Ryotaro
authors.initials: R
authors.lastname: Ito
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Ewa
authors.initials: E
authors.lastname: Ellis
authors.forename: Steven D
authors.initials: SD
authors.lastname: Carson
authors.forename: Shintaro
authors.initials: S
authors.lastname: Sato
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: David
authors.initials: D
authors.lastname: Muirhead
authors.forename: Nalu
authors.initials: N
authors.lastname: Navarro-Alvarez
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Wong
authors.forename: Jayanta
authors.initials: J
authors.lastname: Roy-Chowdhury
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Platt
authors.forename: David F
authors.initials: DF
authors.lastname: Mercer
authors.forename: John D
authors.initials: JD
authors.lastname: Miller
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Strom
authors.forename: Naoya
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Ira J
authors.initials: IJ
authors.lastname: Fox
mesh_terms: D028341:Activins / Q000494:pharmacology; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation* / Q000648:ultrastructure; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D015870:Gene Expression; D053450:Glucocorticoid-Induced TNFR-Related Protein / Q000494:pharmacology; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008854:Microscopy, Electron; D010641:Phenotype; D033581:Stem Cell Transplantation
chemical_list: D053450:Glucocorticoid-Induced TNFR-Related Protein; C422405:activin A; D016222:Fibroblast Growth Factor 2; D028341:Activins; D017228:Hepatocyte Growth Factor; D003907:Dexamethasone