Chunk #16 — Materials and methods — Differentiation of hESCs into cortical interneurons and analysis of ethanol-induced miRNA and mRNA transcriptomic changes by RNA-seq
USA) and 1 μM of purmorphamine (Stemgent, Cambridge, MA, USA), and the medium was changed every other day. For final neuronal differentiation and maturation (Day 19 and after), the cells were cultured in the neuronal maturation medium supplemented with 20 ng/ml of BDNF (R&D Systems, Minneapolis, MN, USA), 200 μM of ascorbic acid (Sigma-Aldrich, St. Louis, MO, USA), and 200 μM of cAMP (Sigma-Aldrich, St. Louis, MO, USA), and the medium was changed every 4 days. After 6 weeks of maturation (totally 62 days in vitro differentiation), the H1 hESC-derived cortical interneurons were characterized by immunostaining (Fig. S3) to confirm the expression of neuronal biomarkers as described in our previous study [33].
