Targeted differentiation of regional ventral neuroprogenitors and related neuronal subtypes from human pluripotent stem cells. Homo sapiens

Purpose: To find the mechanism for the differences in cell fates between the EB (embryoid body) and AD (adherent) culture conditions Methods: hESCs were cultured in different methods, and RNA-seq were generated , in triplicate, using Illumina GAIIx. The sequence reads were analyzed with Weighted gene correlation network analysis (WGCNA) and Gene Ontology (GO) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: We totally found roughly 600 genes which were specifically enriched in both day 4 EB cells and day 4 EB cells treated with SB431542 and LDN193189. Meanwhile, there were around 1000 genes which showed significant higher expression in AD cells Conclusions: complex intracellular cell contexts and extracellular components work together and define the distinct differentiation potencies of the AD and EB cells upon SHH patterning. Overall design: mRNA from hESCs cultured in AD and EB conditions were analysed, in triplicate, using Illumina HiSeq 2000.