GSM3259314: Old mouse aorta scRNA-seq; Mus musculus; RNA-Seq
Isolation of endothelial cells from the aorta was performed by first dissecting the aorta free of adventitia and exposing the endothelium to the surface. After several washes with versine solution (REF), the endothelium was bathed in 1X trypsin solution and incubated for 10min at 37C. After this time and under the dissecting microscope the surface of the endothelium was gently removed with a small scalpel (microfeatherTM) and repeating pipetting to dissociate cell-cell junctions. The trypsin solution with released cells was placed  in a solution of 0.04% BSA and 0.01% FCS. For the generation of single-cell GEMs a suspension of 8700 cells were loaded on a Chromium Single Cell Instrument (10x Genomics) with an estimated targeted cell recovery of ~5000 cells.  Single-cell RNAseq libraries were prepared using the Chromium Single Cell 3' Library & Gel Bead Kit v2 (10x Genomics).