Human neuronal networks on micro-electrode arrays are a highly robust tool to study disease-specific genotype-phenotype correlations invitro.

Britt Mossink,Tamas Kozicz,Chantal Schoenmaker,Dirk Schubert,Hans Van Bokhoven,Anouk H.A Verboven,Monica Frega,Katrin Linda,Eline J.H Van Hugte,Teun M Klein Gunnewiek,Nael Nadif Kasri,Giulia Parodi,Tjitske Kleefstra

doi:10.1016/j.stemcr.2021.07.001

Abstract

SummaryMicro-electrode arrays (MEAs) are increasingly used to characterize neuronal network activity of human induced pluripotent stem cell (hiPSC)-derived neurons. Despite their gain in popularity, MEA recordings from hiPSC-derived neuronal networks are not always used to their full potential in respect to experimental design, execution, and data analysis. Therefore, we benchmarked the robustness of MEA-derived neuronal activity patterns from ten healthy individual control lines, and uncover comparable network phenotypes. To achieve standardization, we provide recommendations on experimental design and analysis. With such standardization, MEAs can be used as a reliable platform to distinguish (disease-specific) network phenotypes. In conclusion, we show that MEAs are a powerful and robust tool to uncover functional neuronal network phenotypes from hiPSC-derived neuronal networks, and provide an important resource to advance the hiPSC field toward the use of MEAs for disease phenotyping and drug discovery.

Highlights

In vitro neuronal models have become an important tool to study the complex communication of healthy and diseased neuronal circuits
With the advancements in human induced pluripotent stem cell technology, the differentiation of human neurons from somatic cells became possible, allowing phenotyping of human neuronal networks. hiPSC-derived neuronal networks on Micro-electrode arrays (MEAs) mimic the activity pattern of rodent neuronal networks, including a stable state of synchronized network bursting, suggesting that they successfully develop into functional neuronal networks (Frega et al, 2019; Fukushima et al, 2016; Kayama et al, 2018; Odawara et al, 2014, 2016; Sasaki et al, 2019)
Excitatory neurons derived from healthy subjects show a comparable phenotype on MEA To investigate if neuronal network activity from hiPSCderived neurogenin 2 (Ngn2)-induced excitatory neurons was reproducible, we performed a meta-analysis on MEA data derived from multiple control lines used in our lab (Frega et al, 2019; Klein Gunnewiek et al, 2020; Mossink et al, 2021)

Summary

Introduction

In vitro neuronal models have become an important tool to study the complex communication of healthy and diseased neuronal circuits. Micro-electrode arrays (MEAs) are cell culture dishes with embedded micro-electrodes that allow noninvasive measurement of neuronal network activity. With the advancements in human induced pluripotent stem cell (hiPSC) technology, the differentiation of human neurons from somatic cells became possible, allowing phenotyping of human neuronal networks. Improvements in MEA analysis software simplified the extraction of parameters that describe the pattern of neuronal activity. These advancements in both human neuronal culturing systems and MEA analysis software contributed to the popularity of MEA technology to study neuronal network phenotypes (Deneault et al, 2019; Frega et al, 2019; Klein Gunnewiek et al, 2020; Wainger et al, 2014)

Results

Discussion

Conclusion