GSM3996888: Thoracic Aorta SMCs, Tsc1+/+  (aka Cntrl); Mus musculus; RNA-Seq
Thoracic aortas were rapidly excised, rinsed in cold PBS, opened longitudinally, sliced into small fragments around 3 mm in size, incubated in 0.5 ml DMEM with 10% FBS, 1.5 mg/ml collagenase A, and 0.5 mg/ml elastase for 60 min at 37°C while the tissue fragments were gently triturated using a pipette every 15 min, passed through a 40 μm filter, incubated with cell-impermeant viability dye for 20 min at 4°C The selected cells were processed for single-cell RNA-seq library preparation using the Chromium™ Single Cell Platform (10x Genomics) as per the manufacturer's protocol: single cells were partitioned into Gel Beads in Emulsion using the Chromium™ system (10x Genomics), followed by cell lysis and barcoded reverse transcription of RNA, cDNA amplification and shearing, and 5' adaptor and sample index attachment. The single-cell RNA-seq libraries were sequenced on a HiSeq 4000 System (Illumina) at the Yale Center for Genome Analysis.