SIX1+PAX3+ Identify a Progenitor for Myogenic Lineage Commitment from hPSCs

To better characterize the transcriptomic profile of hPSC-derived SMPCS, we performed a comparative analysis on the gene signature of FACS-enriched human embryonic (week 9-10), fetal (week 16-20), and hPSC SMPCs and adult SCs using bulk RNA seq. Overall design: We performed 20-30 million reads for each enriched sample which enabled a more in-depth view of the molecular composition of enriched cells. We enriched human SMPCs using the surface markers Lin-ERBB3+NGFR+ (Hicks et al., 2018) and adult SCs using Lin-CD82+NCAM+ (Alexander et al., 2016). Additionally, hPSC SMPCs were depleted for the neural crest marker HNK1, while primary-derived cells were lineage depleted for CD45, CD235A, and CD31. FACS enriched populations, were immediately collected and processed for RNA sequencing (N=3-5 per group). Sequence alignment to the human genome (hg38) using HISAT (Kim et al., 2015) and transcriptome assembly in StringTie (Pertea et al., 2015) were analyzed for differential gene expression (DGE) using DESeq2 with a false discovery rate of q=0.05 (Love et al., 2014).