GSM8061613: Thy, replicate 2, scRNA-seq; Homo sapiens; RNA-Seq
Human thymus samples were obtained from cardiac patients who underwent cardiac surgery. Thymocytes were isolated by density gradient centrifugation using Ficoll-Paque (GE Healthcare Life Science). Single cell suspensions of thymocytes were stained with BV421 conjugated Anti-Human TCRγ/δ or PE conjugated CD1d and APC conjugated MR1 tetramers and enriched with anti-PE and anti-APC microbeads (Miltenyi) according to the manufacturer's instructions. CD3+CD19- cells were identified within the single-cell-resolved lymphocyte gating. CD3+tet+ cells were then sorted and collected in RPMI-1640 with 50% FBS. The sorting purity exceeded 96%. Subsequently, iNKT, MAIT and γδ T cells were combined and processed simultaneously. Single-cell libraries was constructed according to the manufacturer's instructions (10x Genomics). Samples were processed using 10X genomics's Single Cell 5' R2-only chemistry, as per the manufacturer's recommendations.