GSM4278474: Adult WT1; Mus musculus; RNA-Seq
Single cell suspensions were prepared from brain tissue by digestion for 30min at 37ºC with collagenase IV (ThermoFisher), hyaluronidase (Sigma-Aldrich) and DNase I (Sigma-Aldrich). Digested tissue was mechanically disrupted, filtered through 100µm mesh and enriched for leukocytes by centrifugation through 40% Percoll (GE Healthcare). Non-specific binding was blocked using either 2.4G2 supernatant for mouse cells or normal mouse serum for human cells, and dead cells were labelled by fixable viability dye eFluor 780 (ThermoFisher). CD11b+ CD45+ cells were sorted using  BD FACSAria™ III, and resuspended in DPBS 0.04% BSA.Cell viabilityand concentration were assessed by the LUNA-FL Dual Fluorescence Cell Counter. Single-cell libraries were generated using the GemCode Single-Cell Instrument and Single Cell 3′ Library & Gel Bead Kit v2 and Chip Kit (10x Genomics) according to the manufacturer's protocol