Transplantation of stem cell-derived neurons into the damaged neuronal tissue is one of the promising methods for the recovery of neuronal functions. However, little is known about the signal transmission between transplanted stem cell-derived neuronal networks and the host brain tissue during recovery. In this study, we developed microelectrode-array (MEA) substrates with multiple micro-chambers for cell cultures. Each micro-chamber consisted of 4 cell-plating spots with interconnecting conduits fabricated by polydimethylsiloxane (PDMS) lithography. The conduits had the tunneling structures, the height of which was approximately 6 μm, through which only neurites could grow into adjacent cell-plating spots. We cultured mouse cortical neurons in the subset of cell-plating spots, and stem cell-derived neurons were injected into the residual spots 7 days after starting incubation of primary cultures. Intracellular calcium transients as well as electrical activity were detected from both primary cultured-cells. And extracellular electrical activities were recorded from both primary cultured-cells and stem cell-derived neurons in the co-cultures. These results suggested that our MEA-based co-culture system would be a useful tool for in vitro monitoring of transplantation processes of stem cell-derived neurons into host neuronal tissue.
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