GSM6601035: 200054H_sn_pb; Homo sapiens; RNA-Seq
20 mL of heparin anticoagulant PB and BM from SAA patients or HCs were collected to separate mononuclear cells with lymphocyte separation medium (Amersham Bioscience, UK) under sterile conditions. Mononuclear cells were washed with sterile phosphate-buffered saline, The percentage of NK cells was determined by flow cytometry (Calibur, BD Biosciences, USA) using CD3-PerCP (347344, BD Biosciences, USA), CD56-PE (555516, BD Biosciences, USA), or isotype control antibody (BD Biosciences, USA). The remaining cells were suspended in 1ml of sterile magnetic bead sorting buffer and counted. Negative magnetic beads (130-092-657, Miltenyi Biotec, USA) were used to sort NK cells, and the NK cells was washed with sterile phosphate-buffered saline, suspended, and counted. The cell suspension was adjusted to 300-600 cells/μL, and cell viability was analyzed with Countstar Rigel S2. Sequencing analysis was carried out after passing quality inspection. According to the manufacturer's instructions, single-cell RNA-seq libraries were constructed using the Single Cell 3′ Library and Gel Bead Kit v3.1. The libraries were sequenced in an Illumina Novaseq6000 sequencer with a sequencing depth of at least 100,000 reads per cell with a paired-end 150-bp reading strategy (performed by CapitalBio Technology, Beijing).