GSM8005078: C57BL6 mouse aortic arch intima lesser curvature (untreated control); Mus musculus; RNA-Seq
Briefly, aortas were harvested from mice perfused with ice-cold PBS-/- supplemented with 1 μg/mL actinomycin D and 1% heparin. The ascending aorta was isolated and pinned en face in a dish with PBS-/- supplemented with 2 μg/mL actinomycin D. The dissection buffer was washed off, then the tissue was digested for 10 minutes at 37°C with a mixture of Collagenase D and DNAse I in PBS. Following digestion, aortas were washed twice; first with PBS + 2mM EDTA to inactivate the enzymes, followed by PBS alone to remove excess EDTA. Cells from the lesser curvature were gently dislodged and aspirated into PBS-/- (with 0.05% BSA to minimize cell loss and aggregation) with a P10 pipet tip. Libraries where constructed as per the v3 ChromiumTM Single Cell 3' Library Kit manufacturer's protocol.