GSM7999871: AAA_7, scRNAseq; Mus musculus; RNA-Seq
Before taking infrarenal abdominal aorta samples, animals were euthanized by cervical dislocation and injected with 5 ml PBS at the left ventricular apex to perfuse the blood vessels. The infrarenal aorta was removed and crushed. Aortic samples were incubated in hank solution (450 units/ml collagenase I, 125 units/ml XI, 60 units/ml hyaluronidase, and 60 units/ml DNase I dissolved in PBS) at 37 °C for 1 hour. The cell solution was filtered with 70 μm cell mesh and centrifuged to remove lumpy tissues. To isolate CD45-positive immune cells, the cell pellet was resuspended in buffer with CD45 and viability microbeads and magnetically separated. Each group (n=20 AAA, n=50 control) mixes all aortic sample cells. A 10× Chromium Controller (10X Genomics) barcoded the cell solution in Chromium microfluidic chips with 3' chemistry. RNA from barcoded cells was reverse-transcribed and sequencing libraries were created using reagents from a Chromium Single Cell 3' v2 reagent kit (10X Genomics). Illumina's instructions guided sequencing.