On-Chip Agarose Microchamber (AMC) Array Cell-Cultivation System for Topographical Control of Neural Network

Abstract

We have developed a new type of cell-cultivation system that uses agarose microstructures on glass slides and a photo-thermal micro-melting method, thus enabling topographical control of neural network formation during cell cultivation. In this system the nerve cells are immobilized in microstructures, called “microchambers”, within a 50-µm-thick agar layer, forming an agarose microchamber (AMC) array. By using our photo-thermal micro-melting method (called “photo-thermal etching”), the shapes of the microchannels connecting the microchambers can easily be changed, even during cell cultivation, according to the progress of the neural network formation. To demonstrate the capability of this on-chip AMC array culture system for neural cell cultivation and topographical control of network formation, we cultivated PC12 cells in an AMC array. We found that the cells in microchambers made fiber connections through the tunnel-like microchannels after 7 days of cultivation in the presence of NGF. The cells even made fiber connections through new microchannels formed by photo-thermal etching during cultivation. These results suggest that we can form microchannels photo-thermally during cultivation without damaging the cells being cultured in microchambers.