Back-to-back Layered Co-culture on a SiN Porous Membrane for Stable Measurement of Neuronal Extracellular Potentials

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  • Yoshida Satoshi
    Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology
  • Yasuda Takashi
    Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology

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  • ニューロン細胞外電位の安定的計測を目的とするSiN多孔膜を介した階層型共培養
  • ニューロン サイボウ ガイデンイ ノ アンテイテキ ケイソク オ モクテキ ト スル SiN タコウマク オ カイシタ カイソウガタ キョウ バイヨウ

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Abstract

<p>We developed a novel technique for measuring neuronal extracellular potentials in a neuron-astrocyte co-culture system using a SiN (silicon nitride) membrane having microelectrodes and microholes. An 8×8 microelectrode array was formed on one side of a 1 µm-thick SiN membrane, and multiple microholes of 5 µm in diameter were opened through the membrane. Then, neurons were cultured on the microelectrodes after astrocytes were cultured on the back side of the membrane. The neuronal spikes measured in this novel back-to-back co-culture method were compared with those in the conventional method in which both of neurons and astrocytes were co-cultured on the microelectrodes. In the conventional method, formation and enlargement of cellular aggregates made neurons out of touch with microelectrodes, and posed measurement of a small number of spikes. On the other hand, our novel method permitted the firm contact between neurons and microelectrodes and the cell-to-cell interaction through microholes, and therefore neuronal spikes increased drastically with culture time, which led to stable measurement of neuronal extracellular potentials.</p>

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