Next Generation Sequencing Facilitates Quantitative Analysis of Human Pluripotent Stem Cell-Derived Neutrophil-like Cell Transcriptomes

Next-generation sequencing (NGS) has significantly advanced the elucidation of developmental signaling mechanisms that are important for different cell lineage formation from human pluripotent stem cells (hPSCs). We report here the application of RNA-sequencing technology for transcriptome profile of human primary and hPSC-derived neutrophils, and compare to those of hPSCs. Thirteen neutrophil samples from H9 hESCs were performed in IIIumina HiSeq2500. The resulting sequence reads were mapped to human genome (hg19) using HISAT, and the RefSeq transcript levels (RPKMs) were quantified using the python script rpkmforgenes.py. Our RNA-seq data confirmed the stable expression of key neutrophil markers including ITGAM, FUT4, FCGR3A, CEACAM8 and ITGB2. This study shows a detailed analysis of neutrophil transcriptomes generated by RNA-seq technology, providing insight into the mechanisms underlying the differentiation of hPSCs into neutrophils. Overall design: Neutrophil transcriptome profiles of human primary and hPSC-derived neutrophils were generated by RNA-seq technology using IIIumina NextSeq500