Abstract:In recent years, germ-free rat and mice have received extensive attention and application in the field of biomedical research, particularly in the area of gut microbiota. The demand for germ-free rodent experiments has been increasing, posing a challenge for many research institutions to establish qualified germ-free animal populations. The process of breeding germ-free animals for the first generation involves highly specialized and technically challenging work, such as utilizing artificial rearing techniques to feed rodent pups. This review article provides an overview of key aspects of artificial rearing techniques, including nursing methods, preparation of artificial milk, and sterilization methods. It retrospectively summarizes the key technical points and, based on this foundation, offers prospects for further applications of artificial rearing techniques.