Fig. 1

Schematic of the Study Protocol Ex Vivo Fetal Mouse Neurosphere Culture Model with Ethanol Treatment and EV Concentration and Characterization. A NSCs were collected from three separate pregnancies from fetal mouse dorsal telencephalon, segregated by genetic sex, and propagated ex vivo as neurosphere cultures (inset photomicrograph; scale bar, 200 μm). Cultures were treated with control medium or with culture medium containing ethanol (120 or 320 mg/dL), cells were collected and culture-conditioned medium was processed for EV separation and concentration. B Cell culture supernatant was processed by ultracentrifugation protocol for EV separation and concentration (see detailed methods). C Frequency Distribution of EV diameter. Inset depicts Nanosight image of EVs derived from NSCs