Chunk #17 — Materials and methods — Differentiation of hESCs into cortical interneurons and analysis of ethanol-induced miRNA and mRNA transcriptomic changes by RNA-seq
hESC-derived cortical interneurons were then cultured in the neuronal maturation media containing ethanol at a concentration of around 50–100 mM (equivalent to blood alcohol levels of heavy or intoxicated drinkers) for 7 days. The ethanol-containing neuronal maturation medium was changed every other day. After additional 24-h culture without ethanol exposure, the cells were collected for total RNA isolation. Cell treatment experiments (exposed or unexposed to ethanol for 7 days) were performed in duplicate. Extra wells of cells treated with or without ethanol were fixed with 4% paraformaldehyde for cell morphology assay. Ethanol-exposed cells did not show apparent morphological changes (Fig. S4).
