GSM4186885: APOE_ME; Mus musculus; RNA-Seq
The whole aorta was isolated with perivascular adipose tissues and connective tissues removed. Aortic adventitia was then carefully dissected from the medial and intimal layers. The single cell suspensions were then prepared separately from the aortic adventitia and the remaining medial/intimal layers. The adventitia was cut into fine pieces and digested with 2 mg/ml collagenase I  and 2 mg/ml dispase II in Hank's balanced salt solution containing calcium and magnesium for a total 40 min. The remaining aortic media and endothelium were cut open longitudinally and digested with 1 mg/ml papain , 0.5 mg/ml 1,4-Dithioerythritol, 0.156 mg/ml Taurine; 0.25 mg/ml bovine serum albumin in PBS. Single nucleated live cells were sorted into PBS with 0.04% BSA using a flow cytometer and were then analyzed on a 10x Chromium instrument using Chromium Single Cell 3' Reagent Kit v2 chemistry (10X Genomics), following a standard protocol. Library was generated and sequenced on a NovaSeq platform (Illumina) with paired-end 150 bp (PE 150) sequencing strategy. Single-cell RNA-seq