GSM7665531: HC,scRNAseq; Homo; RNA-Seq
A total of 30ml peripheral blood was collected from each sample, and peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation with Ficoll-Paque plus (17144003, Cytiva). After washing twice in PBS, PBMCs were preserved in serum-free cell lyophilization solution (abs9417, absin) and were stored in liquid nitrogen at −196°C until thawing. After PBMC resuscitation, add APC-CD19 (555415, BDBiosciences) and FITC-CD45 (555482, BDBiosciences) and incubate for 30 min. After incubation, centrifuge, wash, and resuspend with an appropriate volume of resuspended buffer and filter through 40 µm cell sieve. Centrifuge at 4°C, discard the supernatant and resuspend the appropriate volume of resuspended buffer. Cells were FACS-sorted by SH800S Cell Sorter (SH800S, SONY) and collected in 15mL tubes with 5mL of media. Data were analyzed using FlowJo v.10 software.