Stimuli‐Responsive Neuronal Networking via Removable Alginate Masks

Abstract

The neuron patterning method has been applied to cultured neuronal networks to achieve better reproducibility and various experimental conditions in brain research. However, as models of dynamic environments, conventional patterned neural networks having an unalterable structure formed by a preprogrammed substrate have limitations, such as replicating the changes in the physical structure of networks in vivo. This study presents a novel culture technique that can control the location of the neuronal cell bodies constituting the network and the connection of the isolated neuronal networks at a desired time point. Alginate hydrogel, which has a cell‐repellent property and is dissolvable by weak chemical treatment, is used as a background masking material for patterning. Axon growth from neural clusters to the background area formed by a microwell‐patterned alginate layer is possible due to removal of the alginate layer, triggering synchronized spontaneous firing of the neuronal clusters. This technique was combined with a microelectrode array system to observe dynamic changes in neuronal activity according to the transformed morphology of the network. Finally, the clustered network structure formed by this technique showed greatly improved spontaneous firing rates and spike amplitudes compared to the network model prepared using an existing uniform structure.

Engineering 3D neuronal networks with directional endogenous neuronal plasticity pathways

“…Furthermore, this is especially true for complex and highly dynamic tissues such as those formed from 3D neuronal networks. While these tissues have been shown to have high degrees of connectivity, synchrony, and bursting behaviors [6][7][8][9] , they consistently show high variability in their resulting firing patterns. Furthermore, while Hebbian plasticity has been shown to be activated in in-vitro cultures to affect the dynamic of neuronal network at a cellular level 10 , achieving control of tissue-wide modulation of the directionality of neuronal activity has yet to be shown.…”

Modulating electrophysiology of motor neural networks via optogenetic stimulation during neurogenesis and synaptogenesis

“…Furthermore, studies have shown varying degrees of emerging phenomena that are correlated to neuronal plastic events that would imply “learning” or “memory storage” in neural circuits 9 – 13 These varying degrees extend from neural mechanisms that regulate potentiation to feedback mechanisms with respect to associations to stimulation. Currently, in-vitro micro electrode array (MEA) systems have aimed to study the effects of stimulation protocols in the resulting network dynamics for the past 30 years 13 – 19 . While some of these studies have induced transient changes in the recorded signals, most have focused on changes of number of action potentials in mature primary neurons and in learning as a result of feedback to strengthen associations to the stimulation patterns.…”

A Versatile Protein and Cell Patterning Method Suitable for Long-Term Neural Cultures

“…Many surface modification techniques have been developed for patterning culture substrates with proteins, polymers, and adhesion promoting molecules. Surface coatings can be patterned with the help of photolithography, , microcontact printing with poly­(dimethylsiloxane) (PDMS) ,− or polyolefin stamps, photobleaching-based cross-linking, masked plasma-etching, UV-based photoscission, laser-interference ablation, polymer brushes, − polymer and biomolecule printing, ,, and many similar methods. − However, despite the wide range of available techniques, they all present important shortcomings.…”