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. 2011 Jul;133(3):350–359. doi: 10.1111/j.1365-2567.2011.03446.x

Figure 4.

Figure 4

Double-negative natural killer (DN NK) cells have developmental potential. Decidual cells in the term trimester were cultured with interleukin-2 (IL-2; 100 U/ml) and/or IL-15 (10 ng/ml) for 7 or 14 days. (a) Representative flow cytometry analysis of the expression of CD27 and CD11b on IL-2-cultured decidual NK (dNK) cells at each time-point. Dot plots were gated on live NK cells using a lymphocyte gate using forward scatter versus side scatter and an NK-cell gate using CD56+ CD3− cells. (b) The frequency of each subset of NK cells at different time-points. Cells were cultured with IL-2. (c) The frequency of each subset of NK cells at different time-points. Cells were cultured with IL-15. (d) The frequency of each NK-cell subset at different time-point. Cells were cultured with IL-2 and IL-15. (e) Gating strategy of sorting DN NK cells. Dot plots were gated on live NK cells using a lymphocyte gate using forward versus side scatter and an NK-cell gate using CD56+ CD3− cells. Then DN NK cells were sorted by gating CD56+ CD3− CD11b− CD27−. The purity of DN NK cells after sorting was > 95%. (f) Representative flow cytometry analysis of the expression of CD27 and CD11b on IL-2-cultured sorted DN NK cells. (g) Representative flow cytometry analysis of the expression of NKG2A on gated IL-2-cultured dNK cells. (h) The frequency of NKG2A+NK cells at each time-point under different culture conditions in (b), (c) and (d). Each result is representative of three experiments.