Reconstruction of three-dimensional osteochondral model by multilayered scaffold-free tissue

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K20181
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: Nagoya University (2021-2022); Kyoto University (2020)
• Principal Investigator: Kim Jeonghyun 名古屋大学, 工学研究科, 助教 (20844591)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 軟骨 / 3次元培養 / スフェロイド / 肥大化 / 軟骨内骨化 / 多細胞 / 計測 / 細胞核

Outline of Final Research Achievements

In this study, we fabricated and evaluated a cartilage scaffold-free tissue (SFT), which is a three-dimensional cell assembly derived from mouse chondroprogenitor ATDC5 cells. After long-term culture without addition of chemical differentiation supplements, hypertrophy of cells within the spheroids was confirmed, and apoptosis was also observed from the center of the spheroids. This is a phenomenon observed in the initial stage of endochondral ossification, and is a result showing the structural effect of the three-dimensional model. In addition, using human mesenchymal stem cell-derived spheroids, it was clarified that the multicellular behavior in the spheroids was greatly changed by chemical differentiation supplements. Moreover, we established an optical clearing technique necessary for cell observation inside the 3D spheroids and further developed a custom-made image analysis pipeline in Python to quantitatively evaluate cell nucleus shape in the spheroids.

Free Research Field

細胞バイオメカニクス

Academic Significance and Societal Importance of the Research Achievements

軟骨を対象としたマウス軟骨前駆細胞(ATDC5)由来３次元細胞組織体scaffold-free tissue(SFT)を作製・評価し、3次元培養法による構造的な効果を示すことができた。また、3次元培養モデルの多細胞挙動計測法や透明化技術を活用した細胞核形状定量計測法を開発し、3次元培養モデルを対象とする新たなin vitro計測技術発展のための学術的意義がある。さらに、これらの技術は移植後生体内におけるin vivoにおける多細胞挙動を予測できる。これらの計測法は、オルガノイドなどの様々な3次元モデルにおける基礎研究及び移植実験による組織工学・再生医療分野に貢献できる社会的意義がある。