Extracellular Vesicles from iPSC-Derived Glial Progenitor Cells Prevent Glutamate-Induced Excitotoxicity by Stabilising Calcium Oscillations and Mitochondrial Depolarization

Neurodegenerative diseases pose a significant challenge to modern medicine. Despite significant advances in neurology, current therapeutic approaches often prove insufficient to treat such disorders. This study investigates the neuroprotective effect of extracellular vesicles derived from glial derivates of human induced pluripotent stem cells. The extra-cellular vesicle’s cargo was characterised by proteomic analysis. They neuroprotective ef-fect was assessed using a model of glutamate excitotoxicity performed on primary culture of cortical neurons. The viability of cortical neurons was estimated using the MTT test and morphometric analyses. A comprehensive methodology was applied to investigate intra-cellular mechanisms, integrating assessments of intracellular calcium concentrations, mitochondrial membrane potential, and targeted inhibition of the PI3K‑Akt pathway. Transcriptomic analysis of cortical neurons was used to validated the role of obtained mechanisms of extracellular vesicle’s neuroprotective effect. The obtaining results demon-strated the improvement of neuronal survival by reducing intracellular calcium levels, and stabilizing mitochondrial membrane potential under glutamate-induced excitotoxi-city via PI3K-Akt signaling pathway activation. Moreover, the vesicles contained proteins that contribute to preventing apoptotic processes, activating regeneration of nervous sys-tem, modulating of calcium ion transport and associated with redox processes. Further transcriptomic analyses of neuroglial cultures treated with EVs showed up-regulation of genes associated with regeneration, inhibition of calcium ion transport, regulation of membrane depolirisation and negative regulation of apoptotic pathways.