2021 Fiscal Year Final Research Report
3D quantitative analysis of substrate transport using enteroids
| Project/Area Number |
19K07185
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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 47060:Clinical pharmacy-related
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| Research Institution | Hokkaido University |
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Principal Investigator |
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | エンテロイド / 画像解析 / トランスポーター / 蛍光観察 |
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| Outline of Final Research Achievements |
The purpose of this study was to establish a substrate transport experimental system using enteloid (small intestine tissue culture system) and apply it to the functional evaluation of transporters. After incorporating the fluorescent substrate into the lumen of the enteloid, the method of automatic contour determination using an image analysis tool was improved. The entire contour could be captured by binarizing the shade of the full-focus bright-field image of the fluorescence microscope, determining the contour by automatic selection, and then applying this contour to the fluorescence image to calculate the fluorescence intensity. At this time, we devised a technique for the microinjection method in which a membrane-impermeable fluorescent reagent is injected into the lumen of the enteloid. This made it possible to analyze the boundary between the lumen and the cell layer while objectively distinguishing it.
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| Free Research Field |
薬物動態学
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| Academic Significance and Societal Importance of the Research Achievements |
消化管吸収機構を明らかにするための既存の方法には，消化管内の生理的状態を保ったままin vitro系で評価する手法や，排出トランスポーターを簡便に評価する実験系が少ないことが問題点であった．小腸から作製されるエンテロイドは，消化管内の生理的状態を維持するため，有用な物質輸送解析のツールになる可能性がある．本研究の成果は，薬物吸収のメカニズムの解明や，薬物の吸収に及ぼす他の薬物あるいは食品成分の影響（相互作用）の解析に有用と考えられる．
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