RNA Sequencing Facilitates Quantitative Analysis of Transcriptomes of human ESCs early hematopoietic differentiation

Purpose:hESCs hematopoietic differentiation recapitulates embryonic hematopoiesis in vivo and occurs through three main stages: mesoderm commitment, hemogenic endothelium (HE) formation and hematopoietic specification. The goal of this study are to increase the understanding of hESCs hematopoietic differentiation process and its regulatory mechanism. Methods: hESC samples were collected through flow cytometry sorting.mRNA profiles of this samples were generated by deep sequencing using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Conclusions: Compared with other three populations, 57 cell surface markers were highly enriched in CD31+CD34+ endothelial cells. Overall design: hESCs hematopoietic differentiation