Fig. 7

Cytotoxicity and apoptosis of sequential seeded co-cultured ovarian cancer spheroids. OvCar8 and Detroit 551, A2780 and Detroit 551 co-cultured spheroids were seeded time-shifted and cultured for 96 h as described in Fig. 2A. Spheroids were treated after 72 h for 24 h with 100 µM cisplatin or PBS. (A,B) Cell toxicity of OvCar8-fibroblast (A) and A2780-fibroblasts (B) spheroids was measured by fluorescence microscopy using CellTox Green 24 h after treatment. Scale bars, 500 μm. The fluorescence signals after treatment were quantified (relative fluorescence units RFU). Quantitative data are means ± SEM, N = 3, t-test * (p < 0.05). (C,D) Viability and caspase activity were measured after 24 h treatmet (C: OvCar8, D: A2780) (relative luminescence units RLU). Quantitative data are means ± SEM, N = 3, t-test * (p < 0.05), ** (p < 0.01), **** (p < 0.0001). (E,F) Viable cells were stained with calcein-AM in green, while death cells appear in red by propidium iodide (E: OvCar8, F: A2780). Scale bars, 200 μm.