biostudies-literatureUnknown9(24)Ng JWKHuman cord blood-derived γδ T cells (CB_γδ) display a highly diverse TCR_γδ repertoire and have a unique subtype composition different from fetal or adult peripheral blood counterparts. We expanded CB_γδ in vitro using an irradiated Epstein-Barr virus-transformed feeder cell-based modified rapid expansion protocol (REP). Single-cell RNA sequencing tracked progressive differentiation of naïve CB_γδ into cells expressing neoantigen-reactive tumor-infiltrating lymphocyte as well as tissue-resident memory precursor-like and antigen-presenting cell-like gene signatures. TCR_γδ clonal tracing revealed a bias toward cytotoxic effector differentiation in a much larger proportion of V_δ2^- clones compared to V_δ2<sup>+<Science advanceseadf3120https://www.ebi.ac.uk/biostudies/studies/S-EPMC10275585biostudies-literatureCord blood-derived V_δ2⁺ and V_δ2^- T cells acquire differential cell state compositions upon in vitro expansion.PMC10275585Ng JWKFan XHwang WYKLim ASTLai JJHTan KWPoon ZLim THGoh YTCheung AMSEaves CJLi SLim TKHGuo DYfalseCord blood-derived V_δ2⁺ and V_δ2^- T cells acquire differential cell state compositions upon in vitro expansion.Human cord blood-derived γδ T cells (CB_γδ) display a highly diverse TCR_γδ repertoire and have a unique subtype composition different from fetal or adult peripheral blood counterparts. We expanded CB_γδ in vitro using an irradiated Epstein-Barr virus-transformed feeder cell-based modified rapid expansion protocol (REP). Single-cell RNA sequencing tracked progressive differentiation of naïve CB_γδ into cells expressing neoantigen-reactive tumor-infiltrating lymphocyte as well as tissue-resident memory precursor-like and antigen-presenting cell-like gene signatures. TCR_γδ clonal tracing revealed a bias toward cytotoxic effector differentiation in a much larger proportion of V_δ2^- clones compared to V_δ2<sup>+<2023-01-01T00:00:00Z2023 Jun2025-07-02T03:04:36.914Z2025-07-02T03:04:36.914ZS-EPMC102755853732734610.1126/sciadv.adf3120