Traditional toxicity testing is facing great challenging as increasing new chemicals and the implementation of“3R”principle． The development of toxicity testing is approaching a pivotal point where expensive and lengthy traditional in vivo animal tests are transformed to in vitro high-throughput alternative methods with quantitative parameters analysis and mechanistic exploration． Animal alternative methods are considered not only for the compliance of “3R”principle，but also for the demands and scientific requirements for the development of toxicology and social economy．The development and application of animal alternative methods have become an important direction of toxicity testing in the twenty-first century，and are gained increasingly wide support and regulatory acceptance with broad prospect and important value．