Whole-transcriptome analysis of endothelial-to-hematopoietic stem cell transition

Hematopoietic stem cells (HSCs) are generated via a natural transdifferentiation process known as endothelial-to-hematopoietic cell transition (EHT). Due to small numbers of embryonal arterial cells undergoing EHT and the paucity of markers to enrich for hemogenic endothelial cells, the genetic program driving HSC emergence is largely unknown. Here, we use a highly sensitive RNAseq method to examine the whole transcriptome of small numbers of enriched aortic HSCs (CD31+cKit+Ly6aGFP+), hemogenic endothelial cells (CD31+cKit-Ly6aGFP+) and endothelial cells (CD31+cKit-Ly6aGFP-). Comparison of mRNA profiles of endothelial cells, hemogenic endothelial cells, and hematopoietic stem cells generated by deep-sequencing of sorted populations from pool of embryos, in triplicate.造血干细胞（Hematopoietic stem cells, HSCs）通过一种被称为内皮细胞向造血细胞转化（Endothelial-to-hematopoietic cell transition, EHT）的自然转分化过程产生。由于发生EHT的胚胎动脉细胞数量稀少，且缺乏可用于富集生血内皮细胞（hemogenic endothelial cells）的标记物，调控HSC生成的遗传程序在很大程度上仍未明确。本研究采用高灵敏度的RNA测序（RNAseq）技术，对少量经富集的主动脉造血干细胞（CD31+cKit+Ly6aGFP+）、生血内皮细胞（CD31+cKit-Ly6aGFP+）及内皮细胞（CD31+cKit-Ly6aGFP-）的全转录组进行分析。我们对混合胚胎来源的分选细胞群体开展深度测序并完成三次生物学重复，进而对内皮细胞、生血内皮细胞与造血干细胞的mRNA表达谱进行比较分析。