Generation of glutamatergic/GABAergic neuronal co-cultures derived from human induced pluripotent stem cells for characterizing E/I balance in vitro

Obtaining mechanistic insights into the disruptions of neuronal excitation and in-hibition (E/I) balance in brain disorders has remained challenging. Here, we pre-sent a protocol for in vitro characterization of E/I balance. Using human induced pluripotent stem cells, we describe the generation of glutamatergic excitatory/ GABAergic inhibitory neuronal co-cultures at defined ratios, followed by analyzing E/I network properties using immunocytochemistry and multi -elec-trode array recording. This approach allows for studying cell-type-specific contri-bution of disease genes to E/I balance in human neurons. For complete details on the use and execution of this protocol, please refer to Mossink et al. (2022)1 and Wang et al. (2022).2

Automated summary

This article presents a protocol for studying the mechanistic insights into disruptions of neuronal excitation and inhibition balance in brain disorders. The authors generated co-cultures of glutamatergic excitatory and GABAergic inhibitory neurons using human induced pluripotent stem cells, and analyzed E/I network properties using immunocytochemistry and multi-electrode array recording. This approach allows for studying the cell-type-specific contribution of disease genes to E/I balance in human neurons.