GSM8697595: human PBMCs from recovered adults_001; Homo sapiens; RNA-Seq
Whole blood was collected in a tube containing ethylene diamine tetraacetic acid (EDTA). PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation with LymphoprepTM solution (AXIS-SHIELD Poc AS, Oslo, Norway) according to the manufacturer's recommendations. The mononuclear cell layer was carefully transferred to a new 15 mL conical tube and washed twice with RPMI 1640 medium (Hyclone, Logan, UT, USA) by centrifuging at 486 × g for 10 min at room temperature. PBMCs were resuspended in RPMI 1640 culture medium containing 10% fetal bovine serum (FBS) (Millipore, Danvers, MA, USA), 100 units/mL penicillin, and 100 μg/mL streptomycin (GIBCO, Grand Island, NY, USA). Single cell RNA (sc-RNA) libraries of PBMCs were prepared according to the guideline from the Chromium Single Cell 5' v3 chemistry (10x Genomics, Cat No. 120237) based on the Drop-seq methodology. The libraries were sequenced using Illumina nova S6000 device by Genergy Biotechnology (Shanghai, China).