01961nas a2200277   4500000000100000000000100001008004100002260001100043653001500054653002400069653002400093653001500117653001400132100002200146700002700168700001900195700002300214700001800237700002200255245012600277856004400403300000800447490000700455520120700462022001401669       2023                            d  c2023/710a3D culture10aalternative methods10ain vitro toxicology10amycotoxins10aspheroids1 aVeronica Zingales1 aMaria Rosaria Esposito1 aNoemi Torriero1 aMercedes Taroncher1 aElisa Cimetta1 aMaría-José Ruiz00aThe Growing Importance of Three-Dimensional Models and Microphysiological Systems in the Assessment of Mycotoxin Toxicity  uhttps://www.mdpi.com/2072-6651/15/7/422  a4220 v153 aCurrent investigations in the field of toxicology mostly rely on 2D cell cultures and animal models. Although well-accepted, the traditional 2D cell-culture approach has evident drawbacks and is distant from the in vivo microenvironment. To overcome these limitations, increasing efforts have been made in the development of alternative models that can better recapitulate the in vivo architecture of tissues and organs. Even though the use of 3D cultures is gaining popularity, there are still open questions on their robustness and standardization. In this review, we discuss the current spheroid culture and organ-on-a-chip techniques as well as the main conceptual and technical considerations for the correct establishment of such models. For each system, the toxicological functional assays are then discussed, highlighting their major advantages, disadvantages, and limitations. Finally, a focus on the applications of 3D cell culture for mycotoxin toxicity assessments is provided. Given the known difficulties in defining the safety ranges of exposure for regulatory agency policies, we are confident that the application of alternative methods may greatly improve the overall risk assessment.  a2072-6651