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Updated: Jan 24, 2026

Design, Surface Treatment, Cellular Plating, and Culturing of Modular Neuronal Networks Composed of Functionally Inter-connected Circuits

Design, Surface Treatment, Cellular Plating, and Culturing of Modular Neuronal Networks Composed of Functionally Inter-connected Circuits

Published on: April 15, 2015

モジュラー神経スフェロイドからの皮質海馬回路の足場フリーアセンブリ：ネットワーク発達とダイナミクスを調査するためのハイスループットプラットフォーム

Ilaria Donati Della Lunga¹, Francesca Callegari², Fabio Poggio³

¹Department of Informatics, Bioengineering, Robotics, Systems Engineering, University of Genoa, Via all'Opera Pia 13, Genoa, Liguria, 1645, ITALY.

|January 22, 2026

Abstract:

Three-dimensional (3D) in vitro models are critical for recapitulating key aspects of neural network development and interregional interactions. We present a scaffold-free modular system based on primary cortical and hippocampal neurospheroids (NSs), which are subsequently coupled to self-assemble into reproducible assembloid-like structures (ASs). Through a multimodal approach, we characterized their morphological, mechanical, metabolic, and functional properties. NSs displayed progressive growth, viability surpassing 2D cultures, and stiffness approaching physiological brain ranges. Immunostaining verified proper neuronal and astrocytic ratios and confirmed a physiologically relevant GABAergic component. Upon coupling, ASs exhibited robust structural integration while maintaining functional modularity. Calcium imaging enabled the investigation of synchronization patterns at modules' interface, while electrophysiology revealed maturation-dependent and configuration-specific emergence of rhythms, a type of activity typically found in vivo. Functional excitation-inhibition balance remained constant throughout development and was pharmacologically modulated successfully. Our platform balances biological relevance and experimental tractability, offering a versatile tool for investigating neural circuit development, network dynamics, and region-specific perturbations in a reproducible and scalable in vitro environment.

キーワード:

アセンブロイド様構造脳リズムカルシウムイメージング皮質海馬マイクロ電極アレイ神経スフェロイド

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