Ascorbic Acid Ameliorates Molecular and Developmental Defects in Human Induced Pluripotent Stem Cell and Cerebral Organoid Models of Fragile X Syndrome

All samples are 50 day old cerebral organoids differentiated from human iPSCs using human the Lancaster protocol (Lancaster et al Nature Protoc, 2014). 4 samples are wild-type (WT) organoids derived from the iPSC line IPSO; 4 samples are Fragile X Syndrome (FX) organoids derived from the iPSC line FX52 in the presence of 15mM HEPES vehicle (Urbach et al, Cell Stem Cell, 2010). 4 samples are Ascorbic Acid treated FX52 cells differentiated into cerebral organoids (FX +AsA). FX +AsA samples were prepared by exposing FX52 iPSCs to 500uM Ascorbic Acid (+15mM HEPES vehicle) for 6 passages (36 days) as iPSCs and then differentiated into cerebral organoids in the presence of Ascorbic Acid throughout the differentiation protocol. By day 50 of the cererbral organoid protocol all organoids were collected for analysis.