In vitro effect of TNF-α concentration on the viability of (A) LNCaP monolayers with freezing to −10°C, 5 min hold time, and (B) MVEC monolayers with freezing to −5°C, 0 min hold time. Cells were treated with TNF-α at concentrations of 0.1, 1, 10, 100, 1000 ng/ml for 4 hours prior to freezing. In vitro effect of TNF-α concentration on DNA fragmentation of (C) LNCaP monolayers with freezing to −10°C, 5 min hold time, and (D) MVEC monolayers with freezing to −5°C, 0 min hold time. Cells were treated with TNF-α at concentrations of 10, 100, 1000 ng/ml for 4 hours prior to freezing. Viability and DNA fragmentation were assessed at 48 hours (LNCaP cells) or 24 hours (MVECs) after freeze/thaw (FT). Values presented are the means ± standard error mean from 3 independent experiments, and control indicates untreated cells. Significant effects of TNF-α compared to control and TNF-α + FT compared to FT alone (*, p<0.05; **, p<0.001).