GSM7766591: 60TM mice, scRNA-seq; Mus musculus; RNA-Seq
All procedures were performed according to the manufacturer's protocol. Briefly, hippocampi were removed and dissociated using a papain-based protocol. After mice were intra-cardially perfused with ice-cold HBSS, the hippocampi were dissected and then digested at 37°C using papain for 20 min. Following mechanical dissociation, the sample was suspended in cold DPBS. The cell suspension was centrifuged at 500×g for 7 min at 4°C. The supernatant was completely aspirated. The cell pellet was resuspended carefully with cold 0.5% BSA/DPBS buffer. After the removal of cell debris. The tube was filled with 0.5% BSA/DPBS buffer. The suspension was passed through a 30um cell filter. After centrifugation at 1000×g for 10 min at 4°C, the supernatant was completely aspirated. The cell pellet was resuspended carefully with cold 0.5% BSA/DPBS buffer. Isolated cell samples were applied to a Chromium system (10x Genomics) according to the manufacturer's instructions. Single cells were captured in droplets with gel beads containing barcoded primers. Libraries were prepared with the Chromium Single Cell 3' Reagent Kit v3 (10x Genomics) and were sequenced with the NextSeq 6000 system (Illumina) at average sequence depths of 63,967 reads/cell (WT mice) and 54,044 reads/cell (60TM mice). The quality of cDNA libraries was assessed using 2100 Expert Bioanalyzer Software (Agilent). scRNA-seq was performed by the Life Science Data Research Center at the University of Tokyo Graduate School of Frontier Sciences.