2020 Fiscal Year Final Research Report
Establishment of long-term culture method of limbal organoids using methylcellulose.
Project/Area Number |
18K09461
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Review Section |
Basic Section 56060:Ophthalmology-related
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Research Institution | Tokyo Dental College |
Principal Investigator |
Higa Kazunari 東京歯科大学, 歯学部, 講師 (60398782)
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Project Period (FY) |
2018-04-01 – 2021-03-31
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Keywords | オルガノイド / 角膜上皮幹細胞 / 細胞治療 |
Outline of Final Research Achievements |
To establish a cultivation method with the limbal epithelial stem cells niche, we performed spheroidal cultivation using methylcellulose, and to investigate whether limbal organoids were engrafted to the limbus, we transplanted human limbal organoids in a rabbit limbal deficiency model.Limbal organoids with a limbal phenotype can be maintained for up to 1 month using methylcellulose. Limbal organoids were successfully engrafted in a rabbit limbal deficiency model. Immunohistochemistry against human nuclei confirmed the presence of organoid-derived cells migrating on to host corneas. Our data suggested that limbal organoids are engrafted to the limbus of a rabbit limbal deficiency model and supply corneal epithelial cells, and that organoids may be an efficient cell source for clinical use.
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Free Research Field |
眼科学
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Academic Significance and Societal Importance of the Research Achievements |
本研究で申請者が確立してきたオルガノイド作成法は他の上皮細胞の分離法に比べ細胞周囲の環境を崩さずに採取できるため、幹細胞-ニッシェの環境を維持していると考えられる。また、異種由来成分が含まれるマトリゲルに変わりすでに化粧品や医薬としても利用されているメチルセルロースを用いてオルガノイド培養法を確立できたことは大きな進歩であり、再生医療新法下で角膜再生への応用が可能な移植法を確立することができたことから、今後、輪部機能不全を伴った難治性の疾患に対する上皮細胞供給の拡大が期待される。
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