There is no unified global standard for regulations and supervisions of food cultures, and the safety evaluation of strains based on the pathogenicity, toxin-producing characteristics and drug resistance, as well as their safe use among consumers are still the focus of regulations in various countries. The safety evaluation techniques and strategies for the potential and emerging risks of food cultures are still in a state of ongoing refinement. With the innovation of technologies, methods and concepts, the application of next-generation risk assessment technology, combined with new clinical (pre-) research methods and new concepts of hierarchical and classification management, have become important means to carry out safety evaluation of food cultures, including genetically modified microorganisms. These advancements have enhanced the scientificity and accuracy of safety evaluation for food cultures, contributing as a significant part of the comprehensive implementation of ‘big food’ concept. This paper systematically reviews new technologies, new methods and new concepts from a global perspective, and their application status of the safety evaluation of food cultures, and provides reference for the supervision of food cultures used in China.