Figure S2.
The expanded NK cells can be refrozen and thawed with minimal loss in viability and cytotoxicity.(A) Viability of NK cells that were frozen in Cryostor CS10 versus FBS plus 10% DMSO. Cells were analyzed immediately after thawing (day 0) or after 2 d of incubation in NK MACS with 1000 U/ml IL-2. Blue, red, and black indicate three different donors (n = 3). (B) Effect of IL-2 supplement and antibody-conjugated beads on cell viability. (C) Cell morphology in NK MACS with different supplementations. Scale bars indicate 200 µm. (D) Reexpansion rate after thawing. (E) In vitro cytotoxicity against K562 cells on day 2 after rethawing. (F) Workflow of NK cell recovery before gene KO. (G) Comparison of cell viability and CD96− cells between the two recovery periods. Data are shown as mean ± SD of three donors (n = 3). Two-tailed Welch's unequal variances t test was used to test for statistical significance. ns, not significant; *, P ≤ 0.05.

The expanded NK cells can be refrozen and thawed with minimal loss in viability and cytotoxicity. (A) Viability of NK cells that were frozen in Cryostor CS10 versus FBS plus 10% DMSO. Cells were analyzed immediately after thawing (day 0) or after 2 d of incubation in NK MACS with 1000 U/ml IL-2. Blue, red, and black indicate three different donors (n = 3). (B) Effect of IL-2 supplement and antibody-conjugated beads on cell viability. (C) Cell morphology in NK MACS with different supplementations. Scale bars indicate 200 µm. (D) Reexpansion rate after thawing. (E) In vitro cytotoxicity against K562 cells on day 2 after rethawing. (F) Workflow of NK cell recovery before gene KO. (G) Comparison of cell viability and CD96− cells between the two recovery periods. Data are shown as mean ± SD of three donors (n = 3). Two-tailed Welch's unequal variances t test was used to test for statistical significance. ns, not significant; *, P ≤ 0.05.

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