Presented Poster at ISSCR 2023

Title

LABEL-FREE FUNCTIONAL CHARACTERIZATION OF HUMAN IPSC-DERIVED NEURONS AT SINGLE-CELL, POPULATION BURST AND SUBCELLULAR RESOLUTION

In this poster you can learn more about how you can:

• Functionally characterize long-term co-cultured human iPSC-derived neurons and astrocytes
• Measure the electrical activity of human iPSC-derived neurons
• Characterize disease models at subcellular resolution

Abstract

In recent years, brain models derived from pluripotent stem cells have become a fundamental tool for studying common neurological disorders, such as epilepsy, Alzheimer’s disease, and Parkinson’s disease. The ability to measure the electrical activity of human iPSC-derived neurons in real time and label-free can provide much needed insights into the complexity of the neuronal networks. Nowadays, combining single cell resolution with high-throughput physiological assays, which can potentially deepen our understanding of subtype-specific neuronal activity, is especially valuable and yet difficult to achieve.

In this study, the MaxTwo System (MaxWell Biosystems, Switzerland), a multi-well high-density (HD)-MEA platform was used. MaxTwo HD-MEA System allows in vitro extracellular recordings of action potentials at different scales, ranging from network through single-neuron to subcellular features. Moreover, we showed the advantages of having an HD-MEA system featuring 26,400 electrodes per well, which are key to increase the statistical power of the data collected from iPSC-derived neurons over multiple days/weeks and to capture the smallest neuronal signals.

Finally, we present the AxonTracking Assay, a tool for automated recording and analysis of individual axonal arbors of many neurons in parallel. The Axon Tracking Assay enables to measure action potential conduction velocity, axonal length, and number of axonal branches. With this unique method, we characterized the function and axonal structure of different iPSC-derived neuronal cell lines.

Our HD-MEA platforms and the extracted metrics, such as firing rate, spike amplitude, and network burst profile among several others, provide an extremely powerful and user-friendly approach for in vitro drug screening and disease modelling.

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