title: Transplantation of hESC-derived hepatocytes protects mice from liver injury.
abstract: BACKGROUND
  Hepatic cell therapy has become a viable alternative to liver transplantation for life-threatening liver diseases. However, the supply of human hepatocytes is limited due to the shortage of suitable donor organs required to isolate high-quality cells. Human pluripotent stem cells reflect a potential renewable source for generating functional hepatocytes. However, most differentiation protocols use undefined matrices or factors of animal origin; as such, the resulting hepatocytes are not Good Manufacturing Practice compliant. Moreover, the preclinical studies employed to assess safety and function of human embryonic stem cell (hESC)-derived hepatocytes are generally limited to immunodeficient mice. In the present study, we evaluate the generation of hepatocytes under defined conditions using a European hESC line (VAL9) which was derived under animal-free conditions. The function capacity of VAL9-derived hepatocytes was assessed by transplantation into mice with acetaminophen-induced acute liver failure, a clinically relevant model.
  
  
  METHODS
  We developed a protocol that successfully differentiates hESCs into bipotent hepatic progenitors under defined conditions, without the use of chromatin modifiers such as dimethyl sulphoxide. These progenitors can be cryopreserved and are able to generate both committed precursors of cholangiocytes and neonate-like hepatocytes.
  
  
  RESULTS
  Thirty days post-differentiation, hESCs expressed hepatocyte-specific markers such as asialoglycoprotein receptor and hepatic nuclear factors including HNF4α. The cells exhibited properties of mature hepatocytes such as urea secretion and UGT1A1 and cytochrome P450 activities. When transplanted into mice with acetaminophen-induced acute liver failure, a model of liver damage, the VAL9-derived hepatocytes efficiently engrafted and proliferated, repopulating up to 10 % of the liver. In these transplanted livers, we observed a significant decrease of liver transaminases and found no evidence of tumourigenicity. Thus, VAL9-derived hepatocytes were able to rescue hepatic function in acetaminophen-treated animals.
  
  
  CONCLUSIONS
  Our study reveals an efficient protocol for differentiating VAL9 hESCs to neonatal hepatocytes which are then able to repopulate livers in vivo without tumour induction. The human hepatocytes are able to rescue liver function in mice with acetaminophen-induced acute toxicity. These results provide proof-of-concept that replacement therapies using hESC-derived hepatocytes are effective for treating liver diseases.
authors.affiliation: INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. laiatolosa@hotmail.com.|Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. laiatolosa@hotmail.com.|Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. laiatolosa@hotmail.com.|DHU Hepatinov, Villejuif, F-94800, France. laiatolosa@hotmail.com.
authors.forename: Laia
authors.initials: L
authors.lastname: Tolosa
authors.affiliation: INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. jerome.caron@inserm.fr.|Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. jerome.caron@inserm.fr.|DHU Hepatinov, Villejuif, F-94800, France. jerome.caron@inserm.fr.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Caron
authors.affiliation: INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. zara.hannoun@inserm.fr.|Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. zara.hannoun@inserm.fr.|DHU Hepatinov, Villejuif, F-94800, France. zara.hannoun@inserm.fr.
authors.forename: Zara
authors.initials: Z
authors.lastname: Hannoun
authors.affiliation: INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. marc-antoni@hotmail.fr.|Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. marc-antoni@hotmail.fr.|DHU Hepatinov, Villejuif, F-94800, France. marc-antoni@hotmail.fr.
authors.forename: Marc
authors.initials: M
authors.lastname: Antoni
authors.affiliation: Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. silvia_lopezcortes@hotmail.com.
authors.forename: Silvia
authors.initials: S
authors.lastname: López
authors.affiliation: CIBERDEM, Centro de Investigacion Prıncipe Felipe, Valencia, S-46012, Spain. dburks@cipf.es.
authors.forename: Deborah
authors.initials: D
authors.lastname: Burks
authors.affiliation: Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. jose.castell@uv.es.|CIBERehd, FIS, Barcelona, S-08036, Spain. jose.castell@uv.es.
authors.forename: Jose Vicente
authors.initials: JV
authors.lastname: Castell
authors.affiliation: INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. anne.weber@inserm.fr.|Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. anne.weber@inserm.fr.|DHU Hepatinov, Villejuif, F-94800, France. anne.weber@inserm.fr.
authors.forename: Anne
authors.initials: A
authors.lastname: Weber
authors.affiliation: Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. gomez_mjo@gva.es.|CIBERehd, FIS, Barcelona, S-08036, Spain. gomez_mjo@gva.es.
authors.forename: Maria-Jose
authors.initials: MJ
authors.lastname: Gomez-Lechon
authors.affiliation: INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. anne.dubart@inserm.fr.|Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. anne.dubart@inserm.fr.|DHU Hepatinov, Villejuif, F-94800, France. anne.dubart@inserm.fr.
authors.forename: Anne
authors.initials: A
authors.lastname: Dubart-Kupperschmitt
mesh_terms: D000082:Acetaminophen / Q000633:toxicity; D000818:Animals; D001659:Biliary Tract / Q000166:cytology; D002454:Cell Differentiation; D002460:Cell Line; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D053595:Embryonic Stem Cells / Q000166:cytology*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D064593:Heterografts; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID
chemical_list: D000082:Acetaminophen