Re-generation of cytotoxic γδT cells with distinctive signatures from human γδT-derived iPSCs

The goal of this study was to compare gene expressions on the single cell level in (i) freshly isolated PBMCs (no stimulation and no sorting), (ii) PBγδT cells; PBMCs were stimulated with HMBPP in vitro and CD3(+) γδTCR(+) cells were sorted and (iii) iγδT cells; differentiated cells from γδT-iPSC clone were stimulated with HMBPP and CD3(+) γδTCR(+) cells were sorted or not sorted. Single-cell RNA sequencing (scRNA-seq) was performed on following seven samples; S1 (freshly isolated PBMCs, Sample Tag01), S2 (CD3+gdTCR+ sorted PB-γδT cells, Sample Tag02), S3 (CD3+gdTCR+ sorted γδ Tcells differentiated from iPSCs, Exp137 d42), S4 (CD3+gdTCR+ sorted γδ Tcells differentiated from iPSCs, Exp138 d41), S5 (CD3+gdTCR+ sorted γδ Tcells differentiated from iPSCs, Exp139, d36), S6 (non-sorted γδ Tcells differentiated from iPSCs, Exp53-5A, d38) and S7 (non-sorted γδ Tcells differentiated from iPSCs, Exp54-5A, d34). Freshly isolated PBMCs, PBγδT cells and iγδT cells were captured and processed using the BD Rhapsody platform, and messenger RNA profiles of 397 genes (BD Rhapsody Immune response panel Hs) were generated by deep sequencing.