The chapter familiarizes the reader with the need to develop, validate and utilize in vitro models to test chemicals for neurotoxic potential. The major advantages and disadvantages of using cell and tissue culture, factors which have stimulated and hampered the promulgation of in vitro models for neurotoxicity testing and recent improvements in tissue culture methodologies are discussed. The rationale of using tissue culture for evaluating neurotoxicants and the unique aspects of culturing nervous tissue will be described. Major considerations for designing screening tests and factors used in selecting a screening battery are discussed. Topics such as parameters in the validation process, choice and quantification of endpoints (i.e., cytotoxic and neurotoxic), cost and technical requirements; choice of test chemicals and other quality control aspects are discussed.