In vitro field potential monitoring on a multi-microelectrode array for the electrophysiological long-term screening of neural stem cell maturation

Abstract

Due to the lack of appropriate cell models as well as automated electrophysiology monitoring technologies, the standardized identification of neurotoxic or protective effects in vitro remains a major problem in today's pharmaceutical ingredient development. Over the past few years, in vivo-like human pluripotent stem cell-derived neuronal networks have turned out to be a promising physiological cell source, if the establishment of robust and time-saving functional maturation strategies based on stable and expandable neural progenitor populations can be achieved. Here, we describe a multi-microelectrode array (MMEA)-based bioelectronics platform that was optimized for long-term electrophysiological activity monitoring of neuronal networks via field potential measurements. Differentiation of small molecule-based neuronal progenitors on MMEAs led to functional neurons within 15 days. More strikingly, these functional neuronal cultures could remain electrophysiologically stable on the MMEAs for more than four weeks. The observed electrophysiological properties correlated with the expression of typical neuron subtype markers and were further validated by specific neurotransmitter applications. With our established monitoring platform, we could show for the first time the long-term stability of the neural stem cell-like progenitor population to differentiate to electrophysiologically active dopaminergic neuronal networks for more than 80 passages. In conclusion, we provide a comprehensive long-term stable field potential monitoring platform based on stem cell-derived human neuronal networks that can be automated and up-scaled for standardized high-content screening applications e.g. in the field of neurotoxic and neuroprotective therapeutics identification.

Literature review and appraisal on alternative neurotoxicity testing methods

“…Moreover, more thorough characterization of the cell system is necessary. Similar to the study by Seidel et al, (2017), statistical analyses are an issue. Here, one well of a 48-well plate is the statistical unit, not electrodes.…”

Advances in Human Stem Cell-Derived Neuronal Cell Culturing and Analysis

“…12.4 Applications of 2D human stem cell-derived neuronal networks Currently, human neuronal networks are increasingly used as in vitro models and their functional measurements give unique information about human neural development, how their properties compare to animal cells, and human diseases and disorders (Canals et al 2015;Odawara et al 2014, Seidel et al 2017Ylä-Outinen et al 2010). Particularly, their usability has been recognized in the areas of (developmental) neurotoxicity, disease modeling and drug screening (Canals et al 2015;Hopkins et al 2015;Johnstone et al 2010;Pamies et al 2016;Tukker et al 2016).…”

In vitro Models for Seizure-Liability Testing Using Induced Pluripotent Stem Cells

“…Multi-electrode array (MEA) systems are increasingly used with iPSC-derived cultures to assess spontaneous electrical activity, synchronous epileptiform bursting activity, drug responses, and network mechanisms, such as long-term potentiation and depression ( Odawara et al, 2014 , 2016a , b ; Frega et al, 2017 ; Ishii et al, 2017 ; Seidel et al, 2017 ; Matsuda et al, 2018 ; Tukker et al, 2018 ). MEAs take advantage of the generation of neuronal ion currents.…”