GSM8173926: tdTomato+ cells, WT, 12wks, scRNA-seq, bio rep 2; Mus musculus; RNA-Seq
LysMCre-RosatdTomato/Apoe−/−mice with or without floxed Srsf3 were fed a high-cholesterol diet for either 12 weeks or 30 weeks. The aortic roots, aortas, and carotid arteries were collected from these mice in RPMI1640 media containing 10% FBS on ice, followed by the digestion with aortic adventitia digestion enzyme solution containing 2.5 mg/ml collagenase II and 1.88 mg/ml Elastase for 5-6 min at 37 °C. After carefully peeling the adventitial layers away, the aortas were cut into 2-5 mm pieces and digested with aorta dissociation enzyme solution containing 0.24 mg/ml DNase I, 5.4 mg/ml collagenase I, 0.24 mg/ml collagenase XI, and 0.22 mg/ml hyaluronidase for 45 min at 37 °C. The resulting suspensions were passed through 70 μm cell strainers and subsequently centrifuged at 400g for 5 min at 4 °C. The pelleted cells were resuspended in pre-cold RPMI1640 media containing 10% FBS and kept on ice for further process. tdTomato+ cells were then FACS sorted using a MoFlo XDP cell sorter for single-cell RNA sequencing. Single-cell RNA-seq libraries were prepared using Chromium Next GEM Single Cell 3' Reagent Kits v.3.1 (10 x Genomics) according to the manufacturer's instructions.