GSM6046152: Mouse-SC-scRNAseq-Young-d6; Mus musculus; RNA-Seq
GSM6046152
Mice were anesthetized with isoflurane and perfused with 1x PBS. The spinal cord was flushed through the spinal canal with PBS. Spinal cord tissues were homogenized with an 18-gauge needle in a protease inhibitor solution created using protease inhibitor cocktail tablets (Roche) per the manufacturer's protocol, centrifuged at 800 g for 5 minutes, and the supernatant was collected and stored at -80°C. Tissue pellet was resuspended in a solution of HBSS with 1 mg/ml collagenase A (Roche) and 1 mg/ml DNase I (Sigma-Aldrich) and incubated at 37°C for 20 minutes. Mononuclear cells were separated from myelin via centrifugation in a 27% Percoll solution (GE Healthcare). Cells were stained for CD45 and sorted via flow cytometry. Mononuclear cells were isolated from 4 spinal cords per sample. Libraries were sequenced on an Illumina Novaseq instrument and counted with CellRanger v3.1.0 using mm10 reference genome GENCODE vM23/Ensembl 98.