02342nas a2200481   4500000000100000000000100001008004100002260001500043653000800058653002800066653002500094653003500119653002600154653001500180100002400195700002200219700002300241700001800264700002000282700001500302700001700317700002000334700001800354700001900372700002700391700001600418700001800434700001600452700001900468700001900487700002400506700002400530700003000554700002100584700002100605700002300626700001500649700001900664245009400683856004700777520102200824022001401846       2025                            d  c2025-02-1310a3Rs10aArtificial intelligence10aHigh content imaging10aImaging-based in vitro methods10aRegulatory toxicology10aValidation1 aMonica Piergiovanni1 aMilena Mennecozzi1 aErio Barale-Thomas1 aDavide Danovi1 aSebastian Dunst1 aDavid Egan1 aAurora Fassi1 aMatthew Hartley1 aPhilipp Kainz1 aKatharina Koch1 aSylvia E. Le Dévédec1 aIris Mangas1 aElena Miranda1 aJo Nyffeler1 aEnrico Pesenti1 aFernanda Ricci1 aChristopher Schmied1 aAlexander Schreiner1 aNadine Stokar-Regenscheit1 aJason R. Swedlow1 aVirginie Uhlmann1 aFredrik C. Wieland1 aAmy Wilson1 aMaurice Whelan00aBridging imaging-based in vitro methods from biomedical research to regulatory toxicology  uhttps://doi.org/10.1007/s00204-024-03922-z3 aImaging technologies are being increasingly used in biomedical research and experimental toxicology to gather morphological and functional information from cellular models. There is a concrete opportunity of incorporating imaging-based in vitro methods in international guidelines to respond to regulatory requirements with human relevant data. To translate these methods from R&D to international regulatory acceptance, the community needs to implement test methods under quality management systems, assess inter-laboratory transferability, and demonstrate data reliability and robustness. This article summarises current challenges associated with image acquisition, image analysis, including artificial intelligence, and data management of imaging-based methods, with examples from the developmental neurotoxicity in vitro battery and phenotypic profiling assays. The article includes considerations on specific needs and potential solutions to design and implement future validation and transferability studies.  a1432-0738