GSM7508837: Atherosclerotic plaque, WT, Animal2, scRNAseq; Mus musculus; RNA-Seq
Single cell suspensions from aortic root, ascending aorta and aortic arch, where most of the aortic plaques are, were prepared by mincing isolated tissues followed by enzymatic digestion with collagenase I (450 U/ml, Cat# LS004196, Worthington), collagenase XI (125 U/ml, Cat# C7657, Sigma-Aldrich), DNase I (60 U/ml, Cat# DN25, Sigma-Aldrich) and hyaluronidase (60 U/ml, Cat# H3506, Sigma-Aldrich) for 1 h at 37°C with agitation (64). Suspensions were then filtered, and dead cells were removed with a Dead Cell Removal Kit (Cat# 480157, Biolegend) to improve cell viability of the samples. Cells were then washed, resuspended in RPMI 1640 + 10% fetal bovine serum and processed further by the Advanced Genomics Core at the University of Michigan. Single cells were partitioned into droplet emulsion using the Chromium Controller (10X Genomics, CA), where cells were lysed and cDNAs were reverse transcribed and barcoded. Amplified cDNAs were used to construct 5' gene expression (GEX) library.All cDNA libraries were sequenced on an Illumina NovaSeq-6000 platform with 150bp paired end reads.