Development of culture method of fish hepatocyte spheroids for in vitro environmental assessment

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K19979
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 63040:Environmental impact assessment-related
• Research Institution: Akita University
• Principal Investigator: Honda Haruka (古賀晴香) 秋田大学, 理工学研究科, 助教 (90756983)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 魚類肝細胞 / スフェロイド / 細胞外マトリクス

Outline of Final Research Achievements

Fish-derived primary cells or cell lines are essential cell sources for environmental assessment and ecotoxicity studies in vitro. In cell-based assays using fish hepatocytes, a monolayer culture method has been used in which cells adhere to the cell culture surface.
In vitro model is the 3D spheroid culture that is well established in mammalian cells because it has been suggested to better reflect in vivo response than monolayer cultures. However, there are few reports about the spheroid culture of fish-derived cells compare with mammalian cells.
The extracellular matrix is an important regulator of cell function. Cells in spheroids interact with intercellular and extracellular matrices. Therefore, this study aims to establish a culture method to improve the function of fish hepatocyte spheroids by adding an extracellular matrix (ECM) during spheroid formation.

Free Research Field

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Academic Significance and Societal Importance of the Research Achievements

年々、使用される化学物質の種類は増加し、複雑な混合物となった環境水が、環境へ悪影響を及ぼすことが懸念されている。排水評価試験の一つに全排水毒性（WET）試験があるが、個体の飼育や維持など煩雑な操作を伴う。WET試験の対象として、簡便に培養や試験が可能な培養細胞の利用が考えられるが、生体の応答性や機能を完全に反映することはできない。本研究の成果によって、長期培養が可能な「in vitro魚類肝臓モデル」を作製することができれば、単層培養では不可能であった慢性毒性や生体蓄積の評価を、in vitroで行うことが可能になる。