Feeder-free directed differentiation of neural crest cells from human embryonic stem cells

Here we compare the transcriptional profile of neural crest cells differentiated on MS5 feeder cells (Lee et al., 2007 Nature Biotechnology) with neural crest cells differentiated in a feeder-free protocol. As controls, neuroepithelial cells (LSB) and wnt-induced neural crest cells were included. Total RNA was isolated at day 18 or 35 of differentiation from FACS-sorted HNK1+/p75+ cells from MS5-induced (MS5-NC), or feeder-free-induced neural crest cells (NPC-NC). Control samples were wnt-induced neural crest cells at day 11 of differentiation (transgenic sox10-expressing human ES cells were used for this sample) and neuroepithelial cells at day 11 (LSB) (Trizol, Sigma). Samples for each group in triplicate were processed for Illumina Bead arrays (Illumina HT-12) by the MSKCC genomics core facility according to the specifications of the manufacturer.