GSM4592584: D_CD45_1; Homo sapiens; RNA-Seq
The dorsal aortae were dissected from human embryos CS15-16 (N=2) then further bisected into ventral and dorsal portions (AoV and AoD respectively). Tissues were dissociated into single cells in 1mg/ml Collagenase-Dispase (Roche) and 0.12 mg/ml of DNase I (Roche) for 35 min in a 37 oC rotating water bath and stained with conjugated antibodies anti-human VE-Cadherin-PE (Beckman Coulter, 5µg/ml), anti-human CD45 –v450 (Clone: HI30, BioLegend, 6µg/ml) and anti-human CD235A – APC (BD Bioscience, 0.2µg/ml) for 1 hour at 4 oC. Dead cells and erythroid cells were excluded by 7AAD and CD235A staining respectively. VC+CD45-, VC+CD45+ and VC-CD45+ populations were sorted using a FACS Aria-II (BD Bioscience) into Eppendorf tubes containing RLT buffer from the RNeasy Micro kit ready for RNA purification. RNA was purified using the RNeasy Micro kit. SMARTer® Stranded Total RNA-Seq Kit v2 -Pico Input Mammalian kit (Takara) for samples 1-12
GEO
GSM4592584