Investigating the pre-lethal cytotoxic effects of bis(2,4-di-tert-butylphenyl)phosphate on Chinese hamster ovary cells using high content analysis

• Bis(2,4-di-tert-butylphenyl)Phosphate can induce subtle pre-lethal cytotoxic effects on Chinese hamster ovary cellular health After 48 h exposure, bis(2,4-di-tert-butylphenyl)Phosphate increased nuclear intensity in cells. Exposure to showed a dose- and time-dependent reduction mitochondrial mass. increase reactive oxygen species time- dependent manner. (bDtBPP) leaches out of polyethylene films used by the biopharmaceutical industry single-use systems (SUS) for culturing drug producing cell lines. Previous studies found bDtBPP (0.025 – 0.110 mg/L) negatively affects (CHO) growth productivity. Less information is known about potential early stages bDtBPP. This study aimed investigate CHO-K1 cells (0.005 - 0.25 μg/ml) at process relevant concentrations following 2, 24 using high content analysis multiple cytotoxicity markers. μg/ml; P ≤ 0.001) intensity. A mass was seen after exposure Reactive 2 μg/ml bDtBPP, 0.05 ( 0.01 0.001). BDtBPP induced health. highlights benefits switching alternative SUS plastics which do not leach may enhance