title: Hybrid sponge comprised of galactosylated chitosan and hyaluronic acid mediates the co-culture of hepatocytes and endothelial cells.
abstract: When constructing an in vitro model of liver tissue to mimic the in vivo liver microenvironment, the major challenge is to preserve and maintain the hepatocyte phenotype. The aim of this study was to develop a novel intelligent hybrid sponge for use in a dense co-culture system designed to simulate the liver microenvironment. We prepared a galactosylated chitosan (GCs)/hyaluronic acid (HA) hybrid sponge using a freeze-drying technique for the co-culture of primary hepatocytes and endothelial cells. Subsequently, we investigated the biocompatibility of the GCs/HA scaffold with primary hepatocytes and endothelial cells in terms of cell attachment, morphology, bioactivity, and maintenance of specific liver functions. The GCs/HA-hybrid sponge demonstrated good biocompatibility not only with primary hepatocytes, but also with endothelial cells. In our model, primary hepatocytes exhibited superior bioactivity and higher levels of liver-specific functions in terms of hepatocyte-specific gene expression, urea production, and testosterone metabolism as compared to a monoculture system. We succeeded in constructing a liver tissue-like model using the GCs/HA-hybrid sponge. Therefore, we anticipate that GCs/HA-hybrid sponges may be a promising matrix for the co-culture of hepatocytes and endothelial cells in liver tissue engineering, and might be employed as a novel co-culture model for applications in toxicology and drug metabolism.
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B51, Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503, Japan.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shang
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B51, Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Tamai
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B51, Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503, Japan.
authors.forename: Ryusei
authors.initials: R
authors.lastname: Ishii
authors.affiliation: Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8525, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Nagaoka
authors.affiliation: Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8525, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yoshida
authors.affiliation: Catalysis Research Center and Graduate School of Life Science, Hokkaido University, Kita-ku, Sapporo 001-0021, Japan.
authors.forename: Masamichi
authors.initials: M
authors.lastname: Ogasawara
authors.affiliation: The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Science, Nankai University, No. 94 Weijin Road, Tianjin 300071, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B51, Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503, Japan. Electronic address: ytagawa@bio.titech.ac.jp.
authors.forename: Yoh-ichi
authors.initials: Y
authors.lastname: Tagawa
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism; D000077192:Adenocarcinoma of Lung; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D018920:Coculture Techniques / Q000379:methods*; D005690:Galactose / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D008175:Lung Neoplasms / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008855:Microscopy, Electron, Scanning; D012333:RNA, Messenger / Q000235:genetics; D017550:Spectroscopy, Fourier Transform Infrared; D013739:Testosterone / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D015415:Biomarkers; D012333:RNA, Messenger; D013739:Testosterone; D006820:Hyaluronic Acid; D048271:Chitosan; D005690:Galactose
keywords: Endothelial cell; Galactosylated chitosan; Hyaluronic acid; Liver; Primary hepatocyte; Tissue