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Updated: Nov 21, 2025
High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents HPHC
Published on: May 10, 2016
Investigating the pre-lethal cytotoxic effects of bis(2,4-di-tert-butylphenyl)phosphate on Chinese hamster ovary
Emma Harper1, Yuling Xie1, Lisa Connolly1
1Institute for Global Food Security, School of Biological Sciences, 19 Chlorine Gardens, BT9 5DL, Queen's University Belfast, Northern Ireland, UK.
Abstract:
Bis(2,4-di-tert-butylphenyl)Phosphate (bDtBPP) leaches out of polyethylene films used by the biopharmaceutical industry in single-use systems (SUS) for the culturing of drug producing cell lines. Previous studies found bDtBPP (0.025 - 0.110 mg/L) negatively affects Chinese hamster ovary (CHO) cell growth and productivity. Less information is known about the potential early stages of subtle pre-lethal cytotoxic effects of bDtBPP. This study aimed to investigate the pre-lethal cytotoxic effects in CHO-K1 cells of bDtBPP (0.005 - 0.25 μg/ml) at process relevant concentrations following 2, 24 and 48 h exposure, using high content analysis to investigate multiple pre-lethal cytotoxicity markers. After 48 h exposure, bDtBPP (0.005 - 0.25 μg/ml; P ≤ 0.001) increased nuclear intensity. A dose- and time-dependent reduction in mitochondrial mass was seen after exposure to bDtBPP. Reactive oxygen species increased after 2 h exposure to 0.25 μg/ml bDtBPP, 24 and 48 h exposure to 0.05 - 0.25 μg/ml bDtBPP (P ≤ 0.01 and P ≤ 0.001). BDtBPP induced subtle pre-lethal cytotoxic effects on CHO-K1 cellular health. This study highlights the cellular health benefits of the biopharmaceutical industry switching to alternative SUS plastics which do not leach bDtBPP, which may enhance CHO-K1 cell productivity.
