Air pollution is a significant threat to human health and it is estimated that globally 8.9 million deaths happen because of it. A link between urban particulate matter exposure and an increased risk of neurodegenerative disease has already been identified. For this reason, I used wild-type hiPSCs to set up a 2D differentiation protocol of about 70 days. Cultures were untreated or treated with two doses of manganese, 1 μM and 20 μM, chosen in relation to scientific literature and after a MTS viability assay. Treatments started from day 40 until the end of differentiation (intermediate time point: day 50). Regarding results, untreated batches showed a correct temporal trend of gene expression of the neural and astrocytic markers according to the protocol. The MTS followed to screen several concentrations of MnCl2 and define those for treatments; both 1 μM and 20 μM did not show a significant reduction in viability after 10 and 30 days of treatment. Immunofluorescence of Cleaved Caspase-3 confirmed the experiment. At glutamatergic and GABAergic synaptic level, I found that MnCl2 alters network connectivity and calcium imaging recordings displayed a reduction of frequency and an increase of amplitude. Moreover, 30 days of 20 μM manganese significantly inhibited Map2 protein production, prompting speculations on cytoskeletal impairment and remodeling and so the impact on the neural network. Regarding other cell types, treatment seems to induce reactive astrogliosis after 10 days. I am currently starting a 3D organoid generation protocol to enhance our understanding of manganese’s neuropathological effects.