Exposure-Based Validation of an In Vitro Gastrulation Model for Developmental Toxicity Assays.

Abstract

Establishment of effective non-animal alternatives for developmental toxicity screening assays is desirable to ensure maternal and fetal health outcomes. Validation of such assays requires a comparison between the in vitro responses to chemical exposures and the in vivo impacts of the corresponding compounds at equivalent concentrations. Here, we investigated how the P19C5 gastrulation model responds to 24 compounds at specific concentrations, some of which are categorized as positive exposures based on previously observed detrimental effects on development in vivo, whereas others are categorized as negative exposures due to lack of effects in vivo. The P19C5 gastrulation model consists of in vitro morphogenesis of mouse stem cells aggregated into embryoid bodies (EBs), which recapitulates growth and axial elongation of early embryos during four days of three-dimensional culture. Adverse impacts of chemical exposures were defined as: death, impaired growth, and altered axial elongation of EBs. Ten out of 17 positive exposures caused adverse impacts on EBs. In contrast, only three out of 17 negative exposures adversely affected EBs, although two of the three diminished viability of somatic cell lines (NIH/3T3, HEK293, and JEG3), suggesting general cytotoxicity. Overall, the study showed that 24 out of 34 exposures impacted EB development in a manner concordant with the in vivo developmental effects. Validation of other alternative assays using the same set of chemical exposures will provide information on the strengths and weaknesses of each assay, and should help determine the most effective ensemble of assays to detect a wide range of developmentally toxic exposures.