Establishment and Analysis of Skin-derived multicellular 3-dimensional in vitro cell models (Organoid)

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K08801
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53050:Dermatology-related
• Research Institution: Keio University
• Principal Investigator: OUCHI TAKESHI 慶應義塾大学, 医学部(信濃町), 専任講師 (30528419)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: ３次元構造体 / オルガノイド / ケラチノサイト

Outline of Final Research Achievements

Mouse skin-derived multicellular 3-dimensional in vitro cell models (Organoid) was established and analyzed. In-vitro generated organoid occur as cysts with a central lumen. Organoid epithelium contains stratum corneum, stratum granulosum, stratum spinosum and basal layer comprising epidermis in vivo. According to the serial observation, it was revealed that aggregated keratinocytes developed into organoids in the presence of cocktails of growth factors. The challenge to overcome was to enable long-term culture. RNA-seq analysis comparing primary and passaging cultures revealed suppression of mTOR signaling pathway upon passaging. The phosphorylation level of Akt was decreased in the passaging culture group, suggesting that down regulated Akt inhibits mTOR pathway. We are currently testing whether SC79, a small molecule that activates Akt, contributes to long-term culture.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

本研究では、マウス皮膚から正常の皮膚（表皮）を試験管内で再現した機能的オルガノイドの培養系を確立した。皮膚の表皮細胞は生体の恒常性の維持に重要な役割を担っており、皮膚の生体外の器官培養系を確立することは、生体外評価モデルの構築として重要な意味を持つ。長期の継代培養が課題であるが、今後、皮膚オルガノイドの長期培養が可能となることで、表皮の恒常性維持、がん化メカニズムを理解するために最適なプラットフォームを提供することが期待される。また遺伝子改変による疾患モデルに応用可能であり、薬剤スクリーニングも容易となり、臨床に貢献しうると考えられる。