Automated high-throughput patch clamp electrophysiology of hiPSC-derived neuronal models

Area of Science:

• Neuroscience
• Stem Cell Biology
• Biophysics

Background:

• Human induced pluripotent stem cells (hiPSCs) offer a powerful model for studying brain disorders.
• Current electrophysiological assays for hiPSC-derived neurons, like manual patch-clamp, lack the throughput for large-scale studies.

Purpose of the Study:

• To establish and validate high-throughput automated patch-clamp (APC) methods for hiPSC-derived neurons.
• To enable efficient investigation of neuronal biophysical properties and drug responses in human models of neurological conditions.

Main Methods:

• Development of dissociation and voltage-clamp recording protocols for hiPSC-derived neurons using the Nanion Syncropatch 384 system.
• Application of APC to neurons derived from 2D cortical differentiation, NGN2 induction, and 3D cortical organoids.
• Integration of fluorescence-activated cell sorting (FACS) with APC for cell-type specific analysis.

Main Results:

• Successful implementation of APC for recording from diverse hiPSC-derived neuronal models.
• Direct comparison of APC with manual patch-clamp, demonstrating comparable accuracy and significantly higher throughput.
• Characterization of voltage-gated sodium channel (VGSC) biophysical properties and demonstration of pharmacological profiling capabilities.

Conclusions:

• Automated patch-clamp provides a transformative, high-throughput solution for assessing electrophysiological properties of hiPSC-derived neurons.
• This technique facilitates compound screening and detailed analysis of neuronal function in various hiPSC models.
• Enables cell-type specific and genetically modified neuronal electrophysiology for advancing brain disorder research.