Abstract
Measurement of the activity of human pluripotent stem cell (hPSC)-derived neuronal networks with microelectrode arrays (MEAs) plays an important role in functional in vitro brain modelling and in neurotoxicological screening. The previously reported hPSC-derived neuronal networks do not, however, exhibit repeatable, stable functional network characteristics similar to rodent cortical cultures, making the interpretation of results difficult. In earlier studies, microtunnels have been used both to control and guide cell growth and amplify the axonal signals of rodent neurons. The aim of the current study was to develop tunnel devices that would facilitate signalling and/or signal detection in entire hPSC-derived neuronal networks containing not only axons, but also somata and dendrites. Therefore, MEA-compatible polydimethylsiloxane (PDMS) tunnel devices with 8 different dimensions were created. The hPSC-derived neurons were cultured in the tunnel devices on MEAs, and the spontaneous electrical activity of the networks was measured for 5 weeks. Although the tunnel devices improved the signal-to-noise ratio only by 1.3-fold at best, they significantly increased the percentage of electrodes detecting neuronal activity (52–100%) compared with the controls (27%). Significantly higher spike and burst counts were also obtained using the tunnel devices. Neuronal networks inside the tunnels were amenable to pharmacological manipulation. The results suggest that tunnel devices encompassing the entire neuronal network can increase the measured spontaneous activity in hPSC-derived neuronal networks on MEAs. Therefore, they can increase the efficiency of functional studies of hPSC-derived networks on MEAs.
Highlights
Analysis of neuronal network activity in vitro is a pivotal part of modern brain disease modelling, neuropharmacological testing, and neurotoxicological screening (Johnstone et al, 2010; Valdivia et al, 2014)
The aim of this study was to develop novel tunnel devices that would facilitate and enhance the detection and analysis of extracellular electrical signals recorded from human pluripotent stem cell (hPSC)-derived neuronal networks
Neurons were cultured on microelectrode arrays (MEAs) covered by PDMS tunnel devices with varying dimensions, and the spontaneous electrical activity was measured for 5 weeks
Summary
Analysis of neuronal network activity in vitro is a pivotal part of modern brain disease modelling, neuropharmacological testing, and neurotoxicological screening (Johnstone et al, 2010; Valdivia et al, 2014). In vitro neuronal networks derived from human pluripotent stem cells (hPSCs) can replace animal-derived models and better predict responses in humans (Cavanaugh et al, 2014; Hunsberger et al, 2015; Pei et al, 2016). Even when the hPSC-derived networks produce robust, measurable activity, the necessary differentiation and functional development can be very slow, taking up to several months (Odawara et al, 2016). It is clear that new approaches are needed to facilitate the analysis of hPSC-derived neuronal network functions
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