Figure 7. NK cells cultured under i-dNK inducing conditions promote HTR trophoblast cell invasion.

a) Representative photomicropgraphs of immortalized extravillous trophoblast HTR-8-svNeo cells in a Matrigel invasion assay. HTR cells were seeded in the upper chamber of matrigel coated migration wells. pNK cells that have been cultured for a week under 1% O₂ in the presence of 2 ng/ml TGFβ1 and 1μM Aza (i-dNK, right panel), or under 21% O₂ and IL-15 as control (left panel) were seeded in the lower chamber. b) i-dNK and control cell data expressed as invasion normalized to control cells (pNK cells from the same donor maintained under 21%O₂ and IL-15 ). Each dot represents one donor. N = 10. * p-value= 0.039. c) Invasion data normalized to media alone in the absence of NK cells for dNK cells from 3 donors , gestational ages 7, 9 and 10 weeks, or i-dNK cells from 4 separate donors (black bars), and reverted i-dNK cells, from the same 4 donors, that were maintained a second week under standard tissue culture basal conditions (21% O₂ +IL-15) (white bars).