We observed numerous instances where EtOH had a differential effect on the transcriptome and methylome of undifferentiated hESCs compared to EBs, which contain differentiating cells encompassing all somatic cell types (Itskovitz-Eldor et al., 2000) including those of neural and hepatic cell lineages. Recent studies have demonstrated inhibitory effects of pharmacologically relevant EtOH concentrations (20-100 mM) on neuronal differentiation of ESCs and specified neural stem cells (NSCs). Specifically, EtOH was shown to inhibit NSC proliferation with dose-dependent increases in apoptosis (Anthony et al., 2008; Fujita et al., 2008; Talens-Visconti et al., 2011) decrease NSC differentiation (Tateno et al., 2004; Zhou et al., 2011) and divert ESC differentiation from neuroectodermal to mesodermal lineage (Ogony et al., 2013; Sanchez-Alvarez et al., 2013). Others utilizing very high EtOH concentrations (300-2000 mM) also observed increased apoptosis of mouse blastocysts (Huang et al., 2007) and increased endodermal differentiation of EBs (Mayshar et al., 2011). Another recent study that focused on differentiation of human hepatocytes from EBs demonstrated that a 48 hr EtOH (20 mM) treatment perturbed the differentiation of progenitor cells into hepatocytes (Pal et al.,
