Construction of neurospheroids via surface modified concave microwells

Developing a three-dimensional (3D) neural tissue model is important to comprehensively understand neural development and neuronal degeneration associated with various neurological disorders such as axonopathy and neuronopathy. Here, a new microplatform suitable for constructing neuronal spheroids (neurospheroids) was developed by modulating cell–surface interactions. The inner surface of a polydimethylsiloxane (PDMS) concave microwell array extensively used in in vitro cell aggregation was modified with typical extracellular matrix (ECM) molecules or carbon nanotubes to control neural spheroid formation. Modulating neuronal cell–ECM interactions could tune 3D intercellular interactions and spheroidal functionality. Neurite outgrowth, a neuronal marker for complex interneuronal signaling, was found to be tightly regulated by cell–ECM interactions in a confined space. Furthermore, amyloid-β (Aβ)-induced axonopathy representing a pathological feature of neurodegenerative diseases in vivo was examined in this study to monitor the degeneration of neurite outgrowth and alteration of neuronal morphology in these neurospheroids. The proposed neural tissue model could be used to study various neurodegenerative diseases in the future.