GSM4943844: mouse_D5_10X; Mus musculus; RNA-Seq
For the D5 dataset, cells isolated from ligated left common carotid arteries of six VSMC lineage-labeled animals (Myh11-EYFP-Ki67/RFP) 5 days after surgery were pooled prior to FACS isolation of EYFP+ cells. 10,000 EYFP+ cells were loaded onto the Chromium system (10x Genomics). Single cell preparation: Carotid arteries were dissected free of perivascular adipose tissue, cut open longitudinally and the endothelium was removed by gentle scraping with a cotton bud. Carotid arteries were then pre-digested in DMEM (Gibco) supplemented with 1 mg/mL Collagenase IV (Invitrogen) and 1 U/mL Elastase (Worthington) for 10 minutes at 37˚C to allow removal of the adventitial layer. The medial layer was transferred to fresh DMEM supplemented with 2.5 mg/mL Collagenase IV (Invitrogen) and 2.5 U/mL Elastase (Worthington) and digested to a single cell suspension at 37 ˚C. A total of 10,000 EYFP+ cells were loaded onto a Chromium system (10x Genomics) and amplified cDNA libraries generated using the 3' Gene Expression v3.0 kit (10x Genomics) were sequenced using 150 cycle paired end protocol (NovaSeq, Illumina).
GEO Accession
GSM4943844