Co-culture and Extracellular Potential Measurement Using Microhole and Microelectrode Arrays

2019 Fiscal Year Final Research Report

• Project/Area Number: 16H03171
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: Kyushu Institute of Technology
• Principal Investigator: YASUDA Takashi 九州工業大学, 大学院生命体工学研究科, 教授 (80270883)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Keywords: マイクロデバイス / 半導体加工 / 微小電極 / 共培養 / 電位計測 / 神経細胞 / アストロサイト

Outline of Final Research Achievements

We developed a technique for co-culturing neurons and astrocytes on both sides of a 1-μm-thick silicon nitride (SiN) membrane which has multiple microholes measuring 3 μm in diameter. By fluorescently staining synapses, we found that the number of synapses formed by a single neuron drastically increases within the first 2 weeks and keeps high values for weeks afterward. This experimental result means that intercellular communication through microholes enhances neuronal activity.
Moreover, we developed a novel microelectrode array device for multichannel measurement of extracellular potentials of neurons which are co-cultured with astrocytes. 64 microelectrodes were formed in an 8×8 matrix on a SiN membrane of 1 μm in thickness having multiple microholes of 6 μm in diameter. Then, neurons and astrocytes were back-to-back co-cultured on both surfaces of the membrane. We succeeded in recording electrical signals caused by spontaneous neuronal activity.

Free Research Field

バイオマイクロデバイス

Academic Significance and Societal Importance of the Research Achievements

本研究の成果により、デバイス上でアストロサイトとの共培養を行うことで神経細胞の活性を長期に渡って維持しながら、神経細胞の電気信号を安定的に計測することが可能になった。この技術は、神経疾患の発症メカニズムの解明や神経疾患治療薬の薬効評価などに応用が可能である。特に、疾患特異的iPS細胞を利用した難治神経疾患の発症機構解明や創薬研究への貢献を期待できる。また、2種類の細胞の組み合わせを、例えば運動ニューロンと骨格筋細胞に変更すれば、筋萎縮性側索硬化症（ALS）の発症メカニズムの解明や治療薬の開発に応用可能であり、本技術は様々な評価系に適用可能な発展性を有している。