Figure 2

The MTD enhances lentiviral transduction of T cells. CD3⁺ enriched T cells were activated for 48 hours using TransAct reagent and transduced with ZsGreen lentiviral vector in the 2M-MTD. (a) Representative experiment for the percent of transduced cells following 2M-MTD or Static transduction of cells at MOI 1.5 for the indicated period of time. ON indicates overnight transduction. MTD reached maximum transduction between 45–90 minutes and was not evaluated overnight for these experiments. Experiment was repeated with similar results. (b) Representative data for the percent of transduced cells following static transduction for 90 minutes or overnight and 2M-MTD transduction for 90 minutes at the indicated MOIs. Experiment was repeated with similar results. (c) Transduction efficiency for repeat experiments following overnight static transduction or 90 minutes MTD transduction at MOI ~1, n = 8 devices with T cells derived from N = 5 different donors, p < 0.05. (d) Percent of T cells recovered from the 2M-MTD with an average of 86%, n = 12 devices with T cells derived from N = 4 different donors. (e) T cell viability relative to time matched static controls immediately following removal from the 2M-MTD and washed in media, n = 19 devices with T cells derived from N = 7 different donors. (f) Cell counts at the indicated days post-transduction for T cells statically transduced for 90 minute or overnight and T cells transduced in the 2M-MTD for 90 minutes. Cells were transduced with 4 different MOI between 1.5 and 15 and the fold expansion for each transduction condition is shown. (g) T cell viability measured by 7AAD and flow cytometry for day 4 post transduction for the same cells in panel f. All error bars indicate standard deviation.