A Protocol for in vitro Culture of highly Active γδ T Cells
The large-scale expansion of γδ T cells in vitro generally requires a long period of time and it is often difficult to maintain cell activity when obtaining high-purity cells.This article establishes an optimized in vitro culture scheme for human γδ T cells without feeder cells and exogenous serum.Based on zoledronic acid(ZOL)and CD3 activator,γδ T cells were expanded from human peripheral blood mononuclear cells(PBMCs),and γδ T cells with a purity of over 90％were obtained on the 6th day of culture.The memory differentiation phenotype of the expanded product was detected using CD62L and CD45RA as markers,with more than 70％being naive T cells and T memory stem cells;detection of CD69 and CD25 indicated that they were in the early to mid-activation stage,and detection of PD-1,LAG-3,and TIM-3 indicated that the degree of exhaustion was low.The expanded product was co-cultured with the Jurkat cell line,and its high anti-tumor cytotoxicity was confirmed by degranulation,IFN-γ secretion,and tumor cell killing rate.The expanded product was infected with lentivirus to confirm that it could accept efficient gene transfection.This study provides highly active γδ T cells that can be used for adoptive cell transplantation,creating new hope for in-depth research and clinical application of tumor immunotherapy.
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