Spatiotemporal regulation of organoid differentiation using a microfluidic probe

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K20658
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: Tohoku University
• Principal Investigator: Nashimoto Yuji 東北大学, 学際科学フロンティア研究所, 助教 (80757617)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Keywords: マイクロ流体プローブ / イオン電流 / 走査型イオンコンダクタンス顕微鏡 / 三次元培養

Outline of Final Research Achievements

The aim of this study was to develop a microfluidic probe to induce and evaluate local differentiation of a tissue model. Using ionic current at the tip, the probe was precisely positioned near a tissue model, a multicellular aggregate. By local injection of a reagent, the nuclei in the model could be locally stained. Next, we investigated whether the probe could be precisely positioned in a microstructure in a tissue model based on ionic current at the probe tip. The position of a microstructure, microvasculature in this study, was successfully detected, and its microstructure was clearly visualized. In addition, a minute amount of cytosol (fL~pL) was precisely collected from a tissue model, by the regulation of the oil-water interface in the probe. In total, based on the ionic current, we developed the methods for fine positioning of the probe, local stimulation, detection of a microstructure, and collection of site-specific cytosols from a tissue model.

Free Research Field

生体医工学

Academic Significance and Societal Importance of the Research Achievements

近年の幹細胞研究は，臓器の一部を模した”ミニ臓器”の作出を可能とし，これらは，“オルガノイド”と呼ばる．しかし，既存のオルガノイドは，幹細胞への，一様な薬剤の添加で，分化が誘導されており，空間的な分化制御は殆ど行われていない．事実，既存のディッシュ培養では，分化が非効率で，バッチ毎のバラつきが課題となっている．本研究で開発した，イオン電流を指標とした探針の位置制御，局所刺激，構造体の検出，回収技術は，局所的な分化誘導，分化評価に応用可能である．探針による局所的な分化制御，分化機能評価という新たな手法の提案により，オルガノイドの分化を人工的に制御する新たな知見が得られることが期待できる．