title: Successful in vivo propagation of factor IX-producing hepatocytes in mice: potential for cell-based therapy in haemophilia B.
abstract: Cell-based therapies using isolated hepatocytes have been proposed to be an attractive application in the treatment of haemophilia B due to the normal production of coagulation factor IX (FIX) in these particular cells. Current cell culture technologies have largely failed to provide adequate isolated hepatocytes, so the present studies were designed to examine a new approach to efficiently proliferate hepatocytes that can retain normal biological function, including the ability to synthesize coagulation factors like FIX. Canine or human primary hepatocytes were transplanted into urokinase-type plasminogen activator-severe combined immunodeficiency (uPA/SCID) transgenic mice. Both donor hepatocytes from canines and humans were found to progressively proliferate in the recipient mouse livers as evidenced by a sharp increase in the circulating blood levels of species-specific albumin, which was correlated with the production and release of canine and human FIX antigen levels into the plasma. Histological examination confirmed that the transplanted canine and human hepatocytes were able to proliferate and occupy >80% of the host livers. In addition, the transplanted hepatocytes demonstrated strong cytoplasmic staining for human FIX, and the secreted coagulation factor IX was found to be haemostatically competent using specific procoagulant assays. In all, the results from the present study indicated that developments based on this technology could provide sufficient FIX-producing hepatocytes for cell-based therapy for haemophilia B.
authors.affiliation: Department of Pediatrics, Nara Medical University, Nara, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Tatsumi
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ohashi
authors.forename: Miho
authors.initials: M
authors.lastname: Kataoka
authors.forename: Chise
authors.initials: C
authors.lastname: Tateno
authors.forename: Masaru
authors.initials: M
authors.lastname: Shibata
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Naka
authors.forename: Midori
authors.initials: M
authors.lastname: Shima
authors.forename: Michiyoshi
authors.initials: M
authors.lastname: Hisanaga
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Kanehiro
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Yoshizato
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Nakajima
authors.forename: Akira
authors.initials: A
authors.lastname: Yoshioka
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D017690:Cell Transplantation / Q000379:methods*; D002478:Cells, Cultured; D002648:Child; D004285:Dogs; D005164:Factor IX / Q000235:genetics / Q000378:metabolism*; D005260:Female; D002836:Hemophilia B / Q000378:metabolism / Q000601:surgery*; D022781:Hepatocytes / Q000378:metabolism / Q000637:transplantation*; D006801:Humans; D007223:Infant; D008099:Liver / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D008822:Mice, Transgenic; D012333:RNA, Messenger / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism; D013997:Time Factors; D014183:Transplantation, Heterologous; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D012709:Serum Albumin; D005164:Factor IX; D014568:Urokinase-Type Plasminogen Activator