Connectivity and network burst properties of in-vitro neuronal networks induced by a clustered structure with alginate hydrogel patterning

Abstract

Modularity is one of the important structural properties that affect information processing and other functionalities of neuronal networks. Researchers have developed in-vitro clustered network models for reproducing the modularity, but it is still challenging to control the segregation and integration of several sub-populations of them. We cultured clustered networks with alginate patterning and collected the electrophysiological signals to investigate the changes in functional properties during the development. We built inter-connected neuronal clusters using alginate micro-patterning with a circular shape on the surface of the micro-electrode array. The neuronal clusters were enabled to be connected at 3 or 10 days-in-vitro (DIV) by removing the barrier. The neuronal signals from different types of networks were collected from 16 to 34 DIV, and functional characteristics were examined. Connectivity and burst motif analysis were carried out to find out the relation between the structure and function of the networks. Neuronal networks with clustered structure showed different activity properties from the random networks along the development. The clustered networks had more short-range connections compared to the random networks. In the network burst motif analysis, the clustered networks showed more various patterns and a slower propagation of the activation patterns. In this study, we successfully cultured neuronal networks with clustered structure, and the structure affected the functional properties. The network model suggested in this study will be a good solution for observing the effect of structure on function during their development.