Regenerative Cartilage made by Fusion of Cartilage Elements derived from Chondrocyte Sheets prepared in Temperature-Responsive Culture Dishes

DOI Web Site 19 References
  • Mori Yoshiyuki
    Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo
  • Kanazawa Sanshiro
    Department of Cartilage & Bone Regeneration (Fujisoft), Graduate School of Medicine, The University of Tokyo
  • Asawa Yukiyo
    Department of Cartilage & Bone Regeneration (Fujisoft), Graduate School of Medicine, The University of Tokyo
  • Sakamoto Tomoaki
    Department of Cartilage & Bone Regeneration (Fujisoft), Graduate School of Medicine, The University of Tokyo
  • Inaki Ryoko
    Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo
  • Okubo Kazumi
    Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo
  • Nagata Satoru
    NAGATA Microtia and Reconstructive Plastic Surgery Clinic
  • Komura Makoto
    Department of Pediatric Surgery, Faculty of Medicine, Saitama Medical University
  • Takato Tsuyoshi
    Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo
  • Hoshi Kazuto
    Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo Department of Cartilage & Bone Regeneration (Fujisoft), Graduate School of Medicine, The University of Tokyo

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Abstract

To establish a large regenerative cartilage without any scaffolds, we cultured chondrocytes on N-Isopropylacrylamide (NIPPAM)-coated dishes with 3 × 3 mm2 grids, producing many cell sheets. We then attempted to have the cell sheets form into pellets, which we term cartilage elements, and have the cartilage elements fuse with each other. Finally, we regenerated the large cartilaginous constructs with some stiffness. The NIPPAM coating on culture dishes, which enables to detach cultured chondrocytes only by the decrease in incubation temperature, was useful for preparing many chondrocyte sheets with a homogenous size. The cell sheets could provide effective matrix production and become spherical to form the cartilage elements. For their fusion, those elements were jammed into an agarose mold and were cultured for 3 weeks. The regenerative constructs made by fusing the cartilage elements derived from human or beagle chondrocytes were subcutaneously transplanted into nude mice and the cell-donor beagles, respectively, showing fair cartilage regeneration 2 months after transplantation. They could also prevent severe foreign-body reactions that often occur when using some scaffolds. Thus, by mass production of homogenous cartilage elements using the NIPPAM-coated dishes and their mutual fusion in the agarose mold, mature cartilage was three-dimensionally regenerated without any scaffolds.

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