GSM8579923: spleen3_5000; Mus musculus; RNA-Seq
The spleens were washed twice with the pre-cooled RPIM1640 (containing 0.04% BSA) and then subjected to scissors for mincing. Subsequently, the tissues were digested in enzymatic hydrolysate containing 3 mg/mL Collagenase I (Gibco, USA) at 37℃ for 1 h. After that, the cell suspensions were twice subjected to a 40μm filter and harvested by centrifugation under 300g for 5 minutes. Cells were then subjected to red blood cell lysis (MACS, Germany) at 4℃ for 10 minutes before enrichment. Finally, the cells were resuspended by 100μl RPIM1640, and cell viability and number were measured using LUNA-FL™ Counter (Logos Biosystems). The single-cell libraries were generated using Chromium Next GEM Single Cell 3ʹ GEM, Library & Gel Bead Kit v3.1 (10× Genomics) and Chip Kit according to the manufacturer's protocol.