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. 2013 Mar 4;8(3):e57922. doi: 10.1371/journal.pone.0057922

Figure 5. Optimizing ex-vivo expansion; Addition of OKT-3.

Figure 5

CD3-depleted cells were expanded in AIM-V or X-VIVO 10 medium with 500 IU/ml IL-2 and one or two rounds of irradiated feeder cells in the presence (“with”) or absence (w/o) of the anti-CD3 antibody OKT3 (30 ng/ml). On the day of initiation OKT3 was added directly to the ready culture comprised of irradiated feeder cells and CD3-depleted cells (10∶1) (“OKT3 directly”) or irradiated feeder were pre-incubated with OKT3 and washed before mixing them with the CD3-depleted cells (“OKT3 pre-incubated”). NK and T cell frequency was defined by flow cytometry analysis in gated live cells. (A) Fold expansion of NK cells after 14 days of expansion; (B) NK cell frequency (%) after 14 days; (C) Fold expansion of NK cells after 21 days; (D) NK cell frequency (%) after 21 days, (E) CD3 T-cell frequency (%) after 21 days; (F) Fold expansion of NK cells after 14 days; (G) Fold expansion of NK cells after 21 days; (H) CD3 T-cell frequency (%) after 21 days.