Stepwise pattern modification of neuronal network during cultivation using photo-thermal etching of agarose architecture

We have developed a new type of on-chip cell cultivation system using an agarose microchamber (AMC) array and a photo-thermal etching method, thus enabling topographical control of neuronal network pattern step-by-step during cell cultivation. By using photo-thermal etching (micro melting) method, the number of microtunnel connecting microchambers can be easily increased, even during cell cultivation, according to the progress of the neuronal network formation. To demonstrate the capability of this system for topographical control of network formation, we cultured hippocampal neurons in this AMC array. We found that the cells in microchambers made fiber connections through microtunnels. Furthermore the cells even made fiber connections through additional microtunnels fabricated during cultivation by photo-thermal etching. The results showed that the photo-thermal etching could be used during cultivation without damaging cells.