Next, we investigated the impact of EtOH exposure on viability of neuronal cells at different lineages. hNSPs, hNPCs, immature neurons (1-week differentiation with no LFP activity), and mature neurons (4-weeks differentiation with high LFP activity) were established as explained above (Fig. 1). Cells were exposed to EtOH at 0, 10, 25, 50, and 75 mM concentrations. To maintain constant ethanol concentrations in cell culture media, we utilized an “anti-evaporation system” as described before27,28. Metabolic activities of the cultures were monitored by an MTT cell viability assay at 6, 24, and 48 h post exposures. As shown in Fig. 2a, cell viability responses of hNSPs were highly sensitive to EtOH as early as 6 h post exposure at 25 mM and higher concentrations. The effect was more dramatic for the 24 and 48 h post exposures and nearly 80–90% of the cells lost their viability at 50 and 75 mm concentrations for 24 and 48 h post exposures. hNPCs also showed reduced metabolic rates and loss of cell viability with a significant decrease at 25, 50, and 75 mM concentrations at
