GSM8586925: C57, replicate 2, scRNAseq; Mus musculus; RNA-Seq
Control skin and skin tumors were excised from mice, minced, and dissociated with a 0.5% collagenase IV (Worthington, LS004188) at 37 °C for 2h. Cells were spun down for 5min at 300g and strained through a 70μm strainer before counting and downstream analysis. Next, cells were stained with CD45-APC (Life technologies, MCD4505) for 30min on ice and then washed with PBS. Live/dead exclusion was performed using 0.1mg/mL DAPI (4',6-diamidino-2'phenylindole dihydrochloride). Then, single cell suspension was used to sort for CD45+ /DAPI- immune cells on a Beckman Coulter XDP100 cell sorter at the University of Colorado Cancer Center Flow Cytometry Shared Resource. The sorted CD45+ cells from two 2323 tumors and two C57 control skins were loaded into a Chromium Single Cell 3' Solution Capture System (10X Genomics). The processed libraries were sequenced on Illumina Novaseq6000 at the Genomics Shared Resource at University of Colorado Anschutz Medical Campus.