OKANO Teruo
   Department   Research Institutes and Facilities, Research Institutes and Facilities
   Position  
Article types Original article
Language English
Peer review Peer reviewed
Title Characterization of chondrocyte sheets prepared using a co-culture method with temperature-responsive culture inserts.
Journal Formal name：Journal of tissue engineering and regenerative medicine
Abbreviation：J Tissue Eng Regen Med
ISSN code：1932-7005(Electronic)1932-6254(Linking)
Domestic / ForeginForegin
Volume, Issue, Page 10(6),pp.486-495
Author and coauthor Kokubo Mami†, Sato Masato*, Yamato Masayuki, Mitani Genya, Kutsuna Toshiharu, Ebihara Goro, Okano Teruo, Mochida Joji
Publication date 2016/06
Summary 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 positiveFN1 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.
DOI 10.1002/term.1764
Document No. 23868865