title: Production of factor VIII by human liver sinusoidal endothelial cells transplanted in immunodeficient uPA mice.
abstract: Liver sinusoidal endothelial cells (LSECs) form a semi-permeable barrier between parenchymal hepatocytes and the blood. LSECs participate in liver metabolism, clearance of pathological agents, immunological responses, architectural maintenance of the liver and synthesis of growth factors and cytokines. LSECs also play an important role in coagulation through the synthesis of Factor VIII (FVIII). Herein, we phenotypically define human LSECs isolated from fetal liver using flow cytometry and immunofluorescence microscopy. Isolated LSECs were cultured and shown to express endothelial markers and markers specific for the LSEC lineage. LSECs were also shown to engraft the liver when human fetal liver cells were transplanted into immunodeficient mice with liver specific expression of the urokinase-type plasminogen activator (uPA) transgene (uPA-NOG mice). Engrafted cells expressed human Factor VIII at levels approaching those found in human plasma. We also demonstrate engraftment of adult LSECs, as well as hepatocytes, transplanted into uPA-NOG mice. We propose that overexpression of uPA provides beneficial conditions for LSEC engraftment due to elevated expression of the angiogenic cytokine, vascular endothelial growth factor. This work provides a detailed characterization of human midgestation LSECs, thereby providing the means for their purification and culture based on their expression of CD14 and CD32 as well as a lack of CD45 expression. The uPA-NOG mouse is shown to be a permissive host for human LSECs and adult hepatocytes, but not fetal hepatoblasts. Thus, these mice provide a useful model system to study these cell types in vivo. Demonstration of human FVIII production by transplanted LSECs encourages further pursuit of LSEC transplantation as a cellular therapy for the treatment of hemophilia A.
authors.affiliation: Blood Systems Research Institute, San Francisco, California, United States of America ; Department of Laboratory Medicine, University of California San Francisco, San Francisco, California, United States of America.
authors.forename: Marina E
authors.initials: ME
authors.lastname: Fomin
authors.forename: Yanchen
authors.initials: Y
authors.lastname: Zhou
authors.forename: Ashley I
authors.initials: AI
authors.lastname: Beyer
authors.forename: Jean
authors.initials: J
authors.lastname: Publicover
authors.forename: Jody L
authors.initials: JL
authors.lastname: Baron
authors.forename: Marcus O
authors.initials: MO
authors.lastname: Muench
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D005169:Factor VIII / Q000096:biosynthesis*; D005333:Fetus / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D006467:Hemophilia A / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D064593:Heterografts; D006801:Humans; D008099:Liver / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics
chemical_list: D015703:Antigens, CD; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C078147:F8 protein, human; D005169:Factor VIII; D014568:Urokinase-Type Plasminogen Activator