GSM7016389: CD45+, after, scRNAseq; Homo sapiens; RNA-Seq
Peripheral blood mononuclear cells (PBMCs) were freshly isolated by density gradient centrifugation from heparinized peripheral blood. Purified PBMCs were sorted into CD45+ leukocytes and CD4+ T cells fractions using a BD FACS Aria III (10,000 cells each, cell viability >98%). CD4+ T cells were stimulated with Phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (500 nM) for 4 h in 1ml cultures. Cells were encapsulated into droplets, and libraries were prepared using Chromium Single Cell 3.1' Reagent Kits v3 according to manufacturer's protocol (10X Genomics). The generated scRNA-seq libraries were sequenced using a 128 cycle (paired-end reads) with a NovaSeq 6000 (Illumina).