GSE161933
Single-cell atlas of major haematopoietic tissues in the mouse embryo
The aim of this experiment was to compare by single-cell RNA sequencing the aorta-gonad-mesonephros (AGM) region and yolk sac (YS), the two major sites of embryonic hematopoietic, between 9.5 and 11.5 days of mouse embryonic development. Overall design: AGM and Yolk Sac from E9.5 and E11.5 C57Bl/6 mouse embryos were isolated and single cell suspensions were generated. Cells were stained with the anti-VE-Cadherin antibody and 7AAD. Cells were analyzed and sorted by flow cytometry. After doublet exclusion, single live 7AAD- cells were bulk sorted into 15ml tubes as VE-CadPos endothelial cells and VE-CadNeg microenvironment cells, and taken for dispensing on ICELL8 chip (Takara). There was two ICELL8 chips in total (one for E9.5 and another for E11.5). Single cells from each population were selected on the ICELL8 machine. Sequencing libraries were generated and were later sequenced together on the Illumina Next Seq 500 sequencer.