Toward Validation of a Human In Vitro Assay for Developmental Toxicity Assessment
Presented at the 9th World Congress on Alternatives and Animal Use in the Life Sciences, August 2014, in Prague, Czech Republic.
Innovative in vitro toxicity screening assays aimed at reducing or replacing the use of animal models are required for the REACH initiative (Europe) and Tox21 initiative (US) to evaluate thousands of chemicals for safety. We have created a predictive, human in vitro pluripotent stem cell based developmental toxicity assay that can reduce costs, animal and compound use, and has potential to increase pharmaceutical and chemical safety. The goal of the current study was to assess the assay’s biomarkers of teratogenicity potential across a broader range of chemicals and migrate the method from a liquid chromatography high resolution mass spectrometry (LC-HRMS) based analysis to a simpler platform. Induced pluripotent stem (iPS) cells were exposed to 8 concentrations of 66 pharmaceutical, environmental and industrial compounds that have been associated with developmental toxicity or considered to be free of developmental toxicity. Spent media was collected and analyzed by LC-HRMS for biomarker discovery and confirmation. The assay was then migrated to a triple quadrupole (QqQ)-LC-MS platform, enabling targeted biomarker analysis and providing for simpler quantification. The results from initial comparisons of the two LC/MS platforms are highly correlated indicating similar assay performance across a broad series of chemical compounds tested.View PDF