Abstract

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Neurological disorders are increasing worldwide, with environmental factors suspected to play a role. To better assess neurotoxicity risks, we developed an advanced in vitro assay using human induced pluripotent stem cell (iPSC)-derived 3D neural cultures composed of cortical neurons and astrocytes. This system exhibits spontaneous calcium oscillations and is optimized for high-throughput screening (HTS). A library of 87 neuroactive and toxic compounds, including drugs, pesticides, and flame retardants, was tested. The platform enabled quantitative ranking of neurotoxic compounds based on their effects on calcium oscillations and cellular health, demonstrating its utility in neurotoxicity assessment.

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Key Learnings
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• iPSC-derived 3D neural cultures provide a physiologically relevant model for neurotoxicity screening.
• Calcium oscillations serve as a functional biomarker for assessing compound effects.
• Multi-parametric analysis enables classification of neurotoxic agents based on their impact on neural activity.
• The platform allows high-throughput evaluation of environmental toxicants and pharmaceutical compounds.

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Methods
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• iPSC-derived cortical neuron/astrocyte co-cultures were utilized in a 384-well plate format.
• Calcium oscillation measurements were performed using high-throughput kinetic fluorescence imaging.
• Cellular and mitochondrial toxicity were assessed via high-content imaging.
• A diverse library of 87 compounds was tested, with effects quantified through multi-parametric data analysis.

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Conclusions
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This study validates iPSC-derived 3D neural cultures as a robust platform for neurotoxicity screening. The system enables high-throughput detection of neurotoxic effects based on calcium oscillations and cellular viability, offering a powerful tool for evaluating environmental and pharmaceutical compounds with potential neurological impact.

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Reference
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Sirenko, O., Parham, F., Dea, S., Sodhi, N., Biesmans, S., Mora-Castilla, S., Ryan, K., Behl, M., Chandy, G., Crittenden, C., Vargas-Hurlston, S., Guicherit, O., Gordon, R., Zanella, F., & Carromeu, C. (2019). Functional and mechanistic neurotoxicity profiling using human iPSC-derived neural 3D cultures. Toxicological Sciences, 167(1), 58–76. https://doi.org/10.1093/toxsci/kfy218