Mycotoxins are naturally occurring toxic secondary metabolites of fungi that may be present in food and feed. Several of these mycotoxins have been associated with human and animal diseases. Fusarium species, found worldwide in cereals and other food types for human and animal consumption, are the most important toxigenic fungi in northern temperate regions. The overall economical loss and the detrimental health effects in humans and animals of mycotoxin contamination are enormous and therefore, rapid screening methods will form an important tool in the protection of humans and animals as well as to minimize economical losses by early detection. An overview of methods for the determination of cytotoxicity and the application of such bioassays to screen solid fungal cultures, cereals, respectively, food/feedstuffs for the presence and toxic potential of Fusarium mycotoxins is presented. Various cell lines including different endpoints of toxicity using vertebrate cells and the predictive value of the in vitro assays are reviewed. Bioassays are compared with existing chemical analytical methods and the possibilities and limitations of such systems are discussed. The review is based on 149 references.