GSM4943846: mouse_D7_10X; Mus musculus; RNA-Seq
To generate the 10X D7 dataset, cells from ligated left common carotid arteries of five VSMC lineage-labeled animals (Myh11-EYFP-Ki67/RFP) 7 days after surgery were pooled. EYFP+ cells were isolated by FACS and enriched for proliferating RFP+EYFP+ cells: a total of 100 EYFP+RFP+ cells were supplemented with 20,000 unselected EYFP+ cells before loading onto a 10X Chromium system. 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 20,000 cells were loaded onto a Chromium system (10x Genomics), amplified cDNA was generated using the 3' Gene Expression v2 kit and sequenced using a paired end protocol (HiSeq 4000, Illumina).
GEO Accession
GSM4943846