Biomedical research is currently in a critical period of transition from traditional animal experiments toward a diversified system combining animal experiments and novel alternative technologies. Conventional animal experiments have historically played a pivotal role in advancing human health. However, with advances in science and technology and evolving ethical considerations, their limitations, including interspecies biological differences, low R&D efficiency, high costs, and ethical controversies, have become increasingly prominent. In this context, emerging alternative technologies such as in vitro biomimetic models (e.g., organoids and organ-on-a-chip systems) and artificial intelligence-driven virtual models have shown great promise in biomedical research, offering advantages such as high physiological and pathological fidelity to human systems and high experimental throughput. Nevertheless, significant challenges remain to be addressed, including the establishment of standardized quality control systems for organoids, optimization of data integration and algorithm interpretability in virtual models, and identification of unaddressed risks. Based on a systematic analysis of the advantages and limitations of traditional animal experiments, this paper explores the value of organoids, organ-on-a-chip systems, and virtual models in addressing the limitations of conventional animal models and the challenges they face in future development, so as to promote the integration and advancement of research technologies based on diverse models and contribute to the advancement of human health.