2021 Fiscal Year Final Research Report
Regenerative potential of human infant small intestinal organoids obtained from surgical specimens
| Project/Area Number |
19K09083
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 55010:General surgery and pediatric surgery-related
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| Research Institution | Juntendo University |
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Principal Investigator |
Koga Hiroyuki 順天堂大学, 大学院医学研究科, 先任准教授 (30468574)
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| Co-Investigator(Kenkyū-buntansha) |
山高 篤行 順天堂大学, 医学部, 教授 (40200703)
松本 有加 順天堂大学, 医学部, 助教 (50813672)
中村 哲也 順天堂大学, 大学院医学研究科, 特任教授 (70265809)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | ヒト腸上皮オルガノイド / 新生児腸疾患 / 腸再生医療 / defined medium |
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| Outline of Final Research Achievements |
We established a protocol for isolating epithelial cells from surgically resected human intestinal specimens from neonates and infants and culturing them as 3D organoids over multiple passages. We found that the organoids can be grown in serum-free media with defined concentrations of Wnt3a and R-spondin 1 which are essential for maintenance of intestinal stem cells. We confirmed that organoids maintained by our protocol are composed of intestinal stem cells and all type of differentiated cells. Furthermore, we also demonstrated that infant small intestinal tissues can generate expandable organoids even after storage for up to 2 days post-resection. This could offer an option of using surgical intestinal specimens from infants to generate organoids when the specimens are expected to be processed within two days, which would broaden the opportunity for the study of human infant intestinal epithelia and their relevance to pediatric diseases.
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| Free Research Field |
小児外科
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| Academic Significance and Societal Importance of the Research Achievements |
既知濃度因子のみを含む無血清培地を用いた培養方法は、オルガノイドを用いた実験の再現性を向上する。また添加因子濃度のカスタマイズが可能であることから細胞分化・増殖などの細胞挙動を開発する研究を可能とする点で学術的意義を持つ。本研究で確立した検体処理・保存方法を用いることで、手術施設で切除した腸組織を別の研究施設に移送してオルガノイドを作製し研究に利用することができる。本研究成果は、新生児・乳児の腸上皮成機構の解明や新生児・乳児期の腸疾患の病態研究の礎の技術になると考える。
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