Impedimetric real-time monitoring of neural pluripotent stem cell differentiation process on microelectrode arrays

Abstract

In today's neurodevelopment and -disease research, human neural stem/progenitor cell-derived networks represent the sole accessible in vitro model possessing a primary phenotype. However, cultivation and moreover, differentiation as well as maturation of human neural stem/progenitor cells are very complex and time-consuming processes. Therefore, techniques for the sensitive non-invasive, real-time monitoring of neuronal differentiation and maturation are highly demanded. Using impedance spectroscopy, the differentiation of several human neural stem/progenitor cell lines was analyzed in detail. After development of an optimum microelectrode array for reliable and sensitive long-term monitoring, distinct cell-dependent impedimetric parameters that could specifically be associated with the progress and quality of neuronal differentiation were identified. Cellular impedance changes correlated well with the temporal regulation of biomolecular progenitor versus mature neural marker expression as well as cellular structure changes accompanying neuronal differentiation. More strikingly, the capability of the impedimetric differentiation monitoring system for the use as a screening tool was demonstrated by applying compounds that are known to promote neuronal differentiation such as the γ-secretase inhibitor DAPT. The non-invasive impedance spectroscopy-based measurement system can be used for sensitive and quantitative monitoring of neuronal differentiation processes. Therefore, this technique could be a very useful tool for quality control of neuronal differentiation and moreover, for neurogenic compound identification and industrial high-content screening demands in the field of safety assessment as well as drug development.

Electrochemical Monitoring of Saos‐2 Cell Differentiation on Pyrolytic Carbon Electrodes

“…In order to analyze more in depth the impedance spectra variations, the impedance spectra were fitted based on an electronic circuit model (Supplementary material S2) . However, comparing cells cultured in GM and differentiation media, the circuit elements used to model the electrode/cell interface showed similar values.…”

Bioimpedance Measurements on Human Neural Stem Cells as a Benchmark for the Development of Smart Mobile Biomedical Applications

“…Numerous studies based on impedance measurements of live biological cells enabled the technique to become widely accepted as a label free, non-invasive and quantitative analytical method to assess cell status. To show some examples, bioimpedance can be used to monitor proliferation [5], apoptosis [6], migration [7], degeneration [8], morphological changes [9] and also (neuronal) differentiation [10].…”

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

“…In this context, we published an actual work that demonstrates the feasibility of using impedance spectroscopy for monitoring and quantifying the initial differentiation process of neuronal differentiation. 7 Nevertheless, for late maturation and functional monitoring of finally differentiated and mature neuronal cultures, the monitoring of neuronal network electrophysiology is crucial, especially in safety assessment to identify the effect of active ingredients on human neuronal cell models. In contrast to traditional technologies such as patch clamp that are invasive and single cell-dependent and hinder scale-up, microelectrode array (MEA)-based field potential recording fulfills the demands of advanced neuronal network electrophysiology characterization.…”