Fig. 8

Vγ2Vδ2 T cells stimulated by pulse zoledronate expand to similar levels using commercial cGMP grade OpTmizer™ media and with larger-scale cultures. a, b Similar purity (a) and yield (b) of Vγ2Vδ2 T cells expanded by pulse zoledronate stimulation in cGMP grade OpTimizer™ culture media as compared with supplemented RPMI 1640 C-media. Vγ2Vδ2 T cells from frozen leukopak PBMC were expanded by pulse zoledronate stimulation with IL-2 for 14 d in either OpTimizer™ culture media or the supplemented RPMI 1640 C-media used in this study. Vγ2Vδ2 T cell numbers were determined by flow cytometric analysis. Representative of two experiments. c, d Expansion of Vγ2Vδ2 T cells by pulse zoledronate culture under larger-scale cultures yielded similar enhancements in yield as noted with small-scale cultures. For larger-scale expansion of Vγ2Vδ2 T cells, leucopak PBMCs were stimulated by either continuous (5 μM) or pulse zoledronate (100 μM) exposure (c) as detailed in Fig. 1 with IL-2 in C-Media in 75 cm² cell culture flasks in triplicate cultures. For (d), Vγ2Vδ2 T cells were expanded by pulse zoledronate stimulation with either IL-2 or IL-15 in C-Media in 75 cm² cell culture flasks in triplicate cultures. After 14 days, Vγ2Vδ2 T cell numbers were determined by flow cytometric analysis. Starting number of Vγ2Vδ2 T cells was 0.158 × 10⁶ for each group. Mean ± SD is shown. **p < 0.01, ***p < 0.001, ****p < 0.0001 using the unpaired t-test