In Vitro Expanded Cord Blood ?d T Cells Acquired Unique Cell State Composition to Mediate Sustained Human AML Suppression [5GEX]

Single cell RNA-seq of cultured gamma-delta T-cells from cord blood were expanded over 2 weeks to identify cell states following in-vitro expansion. Cells were expanded in a feeder co-culture with the EBV-LCL feeder line. After 2 weeks, cells were challenged with the AML cell line K562. Single-cell RNA-sequencing was then performed to identify transcriptomic changes. RNA-seq was paired with single cell TCR sequencing to identify clonotypes mapping to specific cell states. Overall design: Cultured gamma-delta T-cells were sorted with FACS, cultured in the presence of in RPMI + 10% FBS + 170 U/mL recombinant human interleukin-2 (rhIL-2), 25U/ml rhIL-15 as well as 0.7U/ml rhIL-21, with EBV-LCL feeder over a period of 2 weeks and analyzed using scRNAseq before/after antigen challenge with K562.