GSM9041573: Aorta, WT, pooled; Mus musculus; RNA-Seq
After euthanasia, the thoracic aorta and arch were harvested and perivascular fat removed. The aorta was then minced and digested for 45 min at 37˚C in digestion media (DMEM with 1.5 mg/mL Collagenase A, 0.5 mg/mL elastase, 125 U/mL DNAseI, and 10% FBS) with gentle agitation. The digestion was then stopped with additional FBS, washed, and samples were stained with eBioscience Fixable Viability Dye eFluor 780 (Invitrogen) and TotalSeqB Hashtag Oligos (BioLegend) prior to pooling samples from different mice. Single cell suspensions were then generated by FACS sorting: viable tdTom negative and tdTom positive cells were sorted separately and then recombined to enrich tdTom positive cells to ~20% Library prep was performed according to the manufacturer's protocol (10X Genomics Single Cell 3p v4).