Tissue Eng Regen Med.  2016 Aug;13(4):352-363. 10.1007/s13770-016-9065-6.

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

Affiliations
  • 1Graduate School of Biotechnology & Department of Genetic Engineering, College of Life Science, Kyung Hee University Global Campus, Yongin, Korea. ysson@khu.ac.kr
  • 2R&D Institute, MCTT Bio Inc., Seoul, Korea.

Abstract

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.

Keyword

Costal chondrocyte; Bone marrow stromal cell; Cartilage; Chondrogenesis

MeSH Terms

Amnion
Animals
Bone Marrow*
Cartilage
Chondrocytes*
Chondrogenesis
Collagen Type I
Connective Tissue
Humans*
Hyalin*
Hyaline Cartilage*
In Vitro Techniques
Mesenchymal Stromal Cells*
Mice
Phenotype*
Collagen Type I
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