FIG. 2.

Naive T-cell proliferation and activation following direct coculture with ASCs under low O₂ and ambient O₂ culture conditions. Naive CD4⁺ T cells were isolated from peripheral blood of breast cancer patients. ASCs were treated with IFN-γ for 5 days, and then treated with 50 μg/mL mitomycin c to inhibit growth. Naive CD4⁺ T cells were cocultured with IFN-γ-treated ASCs at cell concentrations of 1:100 (1,000 ASCs:100,000 T cells), 1:10 (10,000 ASCs:100,000 T cells), and 1:1 (100,000 ASCs:100,000 T cells) for 3 days. T-cell proliferation was quantified through bromodeoxyuridine (BrdU) incorporation. CD4⁺ T-cell proliferation (%)=(E_Tcell+ASC/E_Tcell)×100, where E_Tcell+ASC=proliferation absorbance measurement for naive T-cell⁺ ASC group, and E_Tcell=proliferation absorbance measurement for T-cell group. (a) Percent (%) T-cell proliferation following coculture at varying ratios. Percent (%) CD25⁺ T cells were evaluated following 1:1 coculture with ASCs by flow cytometry under (b) ambient and (c) low O₂ conditions. (d) Quantification of flow cytometric analyses in panels b and c for CD25 expression of T cells following coculture of ASCs with naive T cells. Three independent sets of experiments were performed for each treatment. Data are reported as mean (μ)±SE. Significance: ***P<0.001.