Developing novel in vitro methods for the risk assessment of developmental and placental toxicants in the environment

During pregnancy, the placenta is critical for the regulation of maternal homeostasis and fetal growth and development. Exposures to environmental chemicals during pregnancy can be detrimental to the health of the placenta and therefore adversely impact maternal and fetal health. Though research on placental-derived developmental toxicity is expanding, testing is limited by the resources required for traditional test methods based on whole animal experimentation. Alternative strategies utilizing in vitro methods are well suited to contribute to more efficient screening of chemical toxicity and identification of biological mechanisms underlying toxicity outcomes. This review aims to summarize methods that can be used to evaluate toxicity resulting from exposures during the prenatal period, with a focus on newer in vitro methods centered on placental toxicity. The following key aspects are reviewed: (i) traditional test methods based on animal developmental toxicity testing, (ii) in vitro methods using monocultures and explant models, as well as more recently developed methods, including co-cultures, placenta-on-a-chip, and 3-dimensional (3D) cell models, (iii) endpoints that are commonly measured using in vitro designs, and (iv) the translation of in vitro methods into chemical evaluations and risk assessment applications. We conclude that findings from in vitro placental models can contribute to the screening of potentially hazardous chemicals, elucidation of chemical mechanism of action, incorporation into adverse outcome pathways, estimation of doses eliciting toxicity, derivation of extrapolation factors, and characterization of overall risk of adverse outcomes, representing key components of chemical regulation in the 21^st century.