Gene expression profiles of EpiSC-derived neural stem cell lines in comparison with EpiSC and embryonic CNS

Epiblast stem cells (EpiSCs) maintained in the medium with activin were freed from activin to elicit neural development, exposed to various strengths of Wnt signals, which may provide the initial stage of anteroposterior regional specificities, and placed in the culture condition to arrest neural development but expand cells as neural stem cells (NSCs) with epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2). Duplicate cultures of original EpiSCs, the neural stem cell lines, XN4, XN1, KN3, KN1, CN1, and CN4, in the order of Wnt signal input levels from lower, were processed for total RNA extraction using TRIzol™ Plus (Thermo Fisher Scientific) and analyzed. Also, the forebrain (FB), midbrain (MB), hindbrain (HB), and cervical spinal cord (SC) tissues of E10.5 mouse embryos were processed for RNA extraction similarly to compare the expression of the central nervous system (CNS) portion landmark genes. Two litters, each with 5~6 embryos, were used as biological duplicates.