title: Characterization of chondrocyte sheets prepared using a co-culture method with temperature-responsive culture inserts.
abstract: Conventional culture methods using temperature-responsive culture dishes require 4-5 weeks to prepare layered chondrocyte sheets that can be used in articular cartilage repair and regeneration. This study investigated whether the use of synovial tissue obtained from the same joint as the chondrocyte nutritive supply source could more quickly facilitate the preparation of chondrocyte sheets. After culturing derived synoviocytes and chondrocytes together (i.e. combined culture or co-culture) on temperature-responsive inserts, chondrocyte growth was assessed and a molecular analysis of the chondrocyte sheets was performed. Transplantable tissue could be obtained more quickly using this method (average 10.5 days). Real-time polymerase chain reaction and immunostaining of the three-layer chondrocyte sheets confirmed the significant expression of genes critical to cartilage maintenance, including type II collagen (COL2), aggrecan-1 and tissue metallopeptidase inhibitor 1. However, the expression of COL1, matrix metalloproteinase 3 (MMP3), MMP13 and A-disintegrin and metalloproteinase with thrombospondin motifs 5 was suppressed. The adhesive factor fibronectin-1 (FN1) was observed in all sheet layers, whereas in sheets generated using conventional preparation methods positive FN1 immunostaining was observed only on the surface of the sheets. The results indicate that synoviocyte co-cultures provide an optimal environment for the preparation of chondrocyte sheets for tissue transplantation and are particularly beneficial for shortening the required culture period. Copyright © 2013 John Wiley & Sons, Ltd.
authors.affiliation: Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Kokubo
authors.affiliation: Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Sato
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo, Women's Medical University, Shinjuku-ku, Tokyo, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamato
authors.affiliation: Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Genya
authors.initials: G
authors.lastname: Mitani
authors.affiliation: Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Kutsuna
authors.affiliation: Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Goro
authors.initials: G
authors.lastname: Ebihara
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo, Women's Medical University, Shinjuku-ku, Tokyo, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
authors.affiliation: Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Joji
authors.initials: J
authors.lastname: Mochida
mesh_terms: D000328:Adult; D000943:Antigens, Differentiation / Q000096:biosynthesis*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D018920:Coculture Techniques; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male
chemical_list: D000943:Antigens, Differentiation
keywords: chondrocyte; co-culture; regeneration; synovial cells; temperature-responsive culture dishes; three-layer chondrocyte sheets