GSM4121360: H8; Mus musculus; RNA-Seq
Blood was rinsed with PBS containing heparin; brains were dissected, collected in ice‐cold PBS, and cut into small pieces. Tissue was digested in 1 ml Accutase (Merck) per brain at 37 °C for 30 min and triturated through 100 μm cell strainers which were rinsed with 10% FCS in PBS. Cells were purified by a linear 40% Percoll (GE Healthcare) centrifugation step at 650 xg without brakes for 25 min and the myelin top layer and supernatant were discarded. Mononuclear cells were resuspended in FACS buffer (1% BSA and 0.1% sodium azide in PBS) and counted. For scRNA-seq cells from brains of 5 12-month-old and 4 24-month-old mice were pooled into 2 separate samples. Around 15,000 CD45highCD8+ single-cells were sorted per sample and encapsulated in droplets using 10X Genomics GemCode Technology kit v2 and processed following manufacturer's specifications. Chromium single cell 3´ v2 (10XGenomics)
GEO Accession
GSM4121360