P18-55 Assessing migration inhibition in neural crest cells using complementary assays

Neural crest cells (NCC) play a pivotal role in early embryogenesis and neurodevelopment, and they express many targets and signaling pathways that may be affected by developmental toxicants. Here, we used the neural crest cell-based migration (cMINC; UKN2) assay to screen for NCC toxicity. Cytotoxicity and cell migration capacity were measured simultaneously, enabling the classification of compounds as being inactive, cytotoxic, borderline active or resulting in a positive activity call in the assay. The interpretation of activity calls in complex cellbased screens is not trivial. To reduce uncertainty, we used a secondary migration assay. Most compounds were successfully confirmed, and the uncertainty concerning borderline hits was reduced. For two exemplary compounds that did not consistently inhibit migration across assays, we demonstrated how such discrepancies can be resolved. We used an additional (tertiary) migration assay that provides detailed information on various aspects of migration, such as cell velocity and directional persistence. On this basis, primary activity calls were validated as DNT-relevant hits.Altogether, this study demonstrates how different migration assays capture distinct aspects of migration inhibition and can complement each other.