4.1 Article

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

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STAR PROTOCOLS
Volume 4, Issue 1, Pages -

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DOI: 10.1016/j.xpro.2022.101967

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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.
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

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