Transcriptome imposed by Notch ligands (Jag1 or Dll4) on ckit+ and ckit- cells from E11.5 AGM. Mus musculus

Hematopoietic Stem Cells (HSC) are originated during embryonic development from endothelial-like cells located in the ventral side of the dorsal aorta around day E10-12 of murine development. This region is called AGM for Aorta/Gonad/Mesonephros refering to the tissues around the hemogenic aorta. Cells that emerge from the endothelium and show hematopoietic traits can be distinguished by the expression of the c-kit receptor and finally acquire the CD45 marker. The effect of Notch activation by each one of the ligands has been tested on c-kit+ and ckit- cells from the endothelium of the E11.5 AGM . Incubations with stromal cell lines exposed the cells to high levels of ligand and induced changes in the transcriptome of these cells. Overall design: AGM regions were obtained from E11.5 embryos by dissection and digested with 0.1% collagenase. Cells were stained with anti-CD31, anti-CD45 and anti-Ter119 antibodies. Sorting of the CD31+CD45-Ter119- population was performed and some cells were separated into ckit+ and ckit- (data previously submited GSE35395). The rest of the CD31+CD45- cells were incubated for 2 hours on OP9 stromal cells expressing ectopic Jagged1 or Delta4 (OP9-Jag1 and OP9-Dll4). After culture with each ligand, cells were separated into CD31+CD45-ckit+ and CD31+CD45-ckit-. Cells were harvested for RNA puriication and microarray analysis.