Effects of ethanol on human-iPSC neurons

This experiment sought to determine the effects of ethanol on induced pluripotent stem cell (iPSC) derived neurons in humans. The iPSC lines were donated by patients with alcohol use disorder (AUD) or without AUD (control) and differentiated through manipulation into neuronal lines. The neuronal lines were cultured for 12 weeks, after which each line was subsequently split into two sister-lines. For the experiment, one sister line was treated by media change with 50mM of ethanol on days 0, 2, 4, and 6 for one week and one sister line was treated by media change with no ethanol on days 0, 2, 4, and 6 for one week. RNA was harvested from both lines on the seventh day with the agent trizol. The harvested RNA was then enriched through rRNA-depletion before 100 cycles of paired amplification. The enriched and amplified RNA-sequences were then read with Illumina HiSeq 2000 and analyzed computationally.