title: Vascularized subcutaneous human liver tissue from engineered hepatocyte/fibroblast sheets in mice.
abstract: Subcutaneous liver tissue engineering is an attractive and minimally invasive approach used to curative treat hepatic failure and inherited liver diseases. However, graft failure occurs frequently due to insufficient infiltration of blood vessels (neoangiogenesis), while the maintenance of hepatocyte phenotype and function requires in vivo development of the complex cellular organization of the hepatic lobule. Here we describe a subcutaneous human liver construction allowing for rapidly vascularized grafts by transplanting engineered cellular sheets consisting of human primary hepatocytes adhered onto a fibroblast layer. The engineered hepatocyte/fibroblast sheets (EHFSs) showed superior expression levels of vascularization-associated growth factors (vascular endothelial growth factor, transforming growth factor beta 1, and hepatocyte growth factor) in vitro. EHFSs developed into vascularized subcutaneous human liver tissues contained glycogen stores, synthesized coagulation factor IX, and showed significantly higher synthesis rates of liver-specific proteins (albumin and alpha 1 anti-trypsin) in vivo than tissues from hepatocyte-only sheets. The present study describes a new approach for vascularized human liver organogenesis under mouse skin. This approach could prove valuable for establishing novel cell therapies for liver diseases.
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan. Electronic address: y.sakai.bioeng@gmail.com.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sakai
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Kosho
authors.initials: K
authors.lastname: Yamanouchi
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ohashi
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Koike
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Utoh
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Hideko
authors.initials: H
authors.lastname: Hasegawa
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Izumi
authors.initials: I
authors.lastname: Muraoka
authors.affiliation: Central Electron Microscope Laboratory, Nagasaki University School of Medicine, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Suematsu
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Soyama
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Hidaka
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Mitsuhisa
authors.initials: M
authors.lastname: Takatsuki
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Kuroki
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan. Electronic address: sueguchi@nagasaki-u.ac.jp.
authors.forename: Susumu
authors.initials: S
authors.lastname: Eguchi
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D016513:Mice, SCID; D008875:Middle Aged; D040521:Subcutaneous Tissue / Q000098:blood supply* / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins
keywords: Cell sheet; Fibroblast; Hepatocyte transplantation; Human primary hepatocyte; Tissue engineering; Vascularization