title: Repopulation of athymic mouse liver by cryopreserved early human fetal hepatoblasts.
abstract: Transplantation of hepatocytes is a promising alternative to liver transplantation for the treatment of severe liver diseases. However, this approach is hampered by the shortage of donor organs and intrinsic limitations of adult hepatocytes. To investigate whether most of the hurdles faced with adult hepatocytes could be surmounted by the use of human fetal hepatoblasts, we have developed a method to isolate, transduce, and cryopreserve hepatoblasts from human livers at an early stage of development (11-13 weeks of gestation). Cells were characterized in vitro for expression of specific markers, and in vivo for their proliferation and differentiation potential after transplantation into athymic mice. Most of the cells (80-90%) harbored a bipotent phenotype, expressing cytokeratins 8/18, albumin, and CK19. They proliferated spontaneously in culture and were efficiently transduced by a beta-galactosidase-expressing retrovirus (90%). After transplantation, cryopreserved cells engrafted into the liver of athymic mice and proliferated, resulting in up to 10% repopulation. Engrafted cells expressed markers of differentiated adult hepatocytes including albumin, alpha1-antitrypsin, cytochrome P450 3A4, and alpha-glutathione-S-transferase. When retrovirally transduced before transplantation they expressed the transgene in vivo. In summary, early human fetal hepatoblasts engraft, proliferate, and mature in athymic mouse liver, without conditioning the donor.
authors.affiliation: Equipe Mixte INSERM 00-20, Laboratoire de Transfert de Gènes dans le Foie: Applications Thérapeutiques, Hôpital Bicêtre, 94270 Le Kremlin Bicêtre and University Paris XI, France. dominique.mahieu@kb.inserm.fr
authors.forename: Dominique
authors.initials: D
authors.lastname: Mahieu-Caputo
authors.forename: Jean-Etienne
authors.initials: JE
authors.lastname: Allain
authors.forename: Julie
authors.initials: J
authors.lastname: Branger
authors.forename: Aurore
authors.initials: A
authors.lastname: Coulomb
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Delgado
authors.forename: Marion
authors.initials: M
authors.lastname: Andreoletti
authors.forename: Sylvie
authors.initials: S
authors.lastname: Mainot
authors.forename: René
authors.initials: R
authors.lastname: Frydman
authors.forename: Philippe
authors.initials: P
authors.lastname: Leboulch
authors.forename: James P
authors.initials: JP
authors.lastname: Di Santo
authors.forename: Frédérique
authors.initials: F
authors.lastname: Capron
authors.forename: Anne
authors.initials: A
authors.lastname: Weber
mesh_terms: D000418:Albumins / Q000032:analysis / Q000235:genetics; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002455:Cell Division; D015925:Cryopreservation; D016332:Fetal Tissue Transplantation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D012190:Retroviridae / Q000235:genetics; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous
chemical_list: D000418:Albumins; D015415:Biomarkers; D049452:Green Fluorescent Proteins; D007633:Keratins