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

Abstract

Summary: Obtaining mechanistic insights into the disruptions of neuronal excitation and inhibition (E/I) balance in brain disorders has remained challenging. Here, we present 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-electrode array recording. This approach allows for studying cell-type-specific contribution 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cell culture
• Microscopy
• Neuroscience
• Stem Cells
• Cell Differentiation