Disc type hyaline cartilage reconstruction using 3D cell sheet culture of human bone marrow stromal cells and human costal chondrocytes and maintenance of its shape and phenotype after transplantation
In this study, we developed the disc type bio cartilage reconstruction strategies for transplantable hyaline cartilage for reconstructive surgery using 3D cell sheet culture of human bone marrow stromal cells and human costal chondrocytes. We compared chondrogenesis efficiency between different chondrogenic induction methods such as micromass culture, pellet culture, and 3D cell sheet culture. Among them, the 3D cell sheet culture resulted in the best chondrogenesis with the disc type bio cartilage >12 mm diameter in size) in vitro, but sometimes spontaneous curling and contraction of 3D cell sheet culture resulted in the formation of bead type cartilage, which was prevented by type I collagen coating or by culturing on amniotic membrane. Previously, it was reported that tissue engineered cartilage reconstructed in vitro does not maintain its cartilage phenotype after transplantation but tends to transform to other tissue type such as bone or connective tissue. However, the disc type bio cartilage of 3D cell sheet culture maintained its hyaline cartilage phenotype even after exposure to the osteogenic induction condition in vitro for 3 weeks or after the transplantation for 4 weeks in mouse subcutaneous. Collectively, the disc type bio cartilage with 12 mm diameter can be reproducibly reconstructed by the 3D cell sheet culture, whose hyaline cartilage phenotype and shape can be maintained under the osteogenic induction condition as well as after the transplantation. This disc type bio cartilage can be proposed for the application to reconstructive surgery and repair of disc type cartilage such as mandibular cartilage and digits.
Tissue Engineering and Regenerative Medicine