GSM8135736: IL-12/18 Adult F1 [scRNA-seq]; Homo sapiens; RNA-Seq
CD8+ cells were magnetically enriched by positive selection (anti-human CD8 microbeads; Miltenyi Biotech) from rested PBMCs or CBMCs. 2x105 cells were plated per well in round-bottomed 96 well plates. Cells were incubated in RP-10 supplemented with recombinant human IL-2, recombinant human IL-12p70 and recombinant human IL-18 at 10ng/ml for 18-22 hours. Live CD235a- CD4- CD8+ cells were sorted into 0.04% BSA in PBS. Cells were counted and 3-8k cells per sample were loaded in the Chromium controller for the formation of gel bead-in-emulsions (GEMs) following the manufacturer's instructions (10X Genomics). The single-cell RNA-seq libraries were prepared using Chromium Single Cell 3′ Reagent Kit v3 following the accompanied protocol and were sequenced on Illumina NextSeq500.