The study of cancer heavily relies on preclinical models to gain insights into disease mechanisms and develop effective therapies. Murine and rat models are particularly noteworthy due to their genetic similarity to humans and cost-effectiveness. In this special issue, we explore advancements in using these models with a special view to therapeutic targets and drug efficacy and follow-up therapeutic interventions. Furthermore, immunocompetent models shed light on the interaction between the immune system and cancer, thus paving the way for immunotherapy, while xenografts can be of interest in other type of investigation and are usually preludes for clinical investigation. This special issue covers the diverse range of models employed in cancer research and their valuable contributions to advancing our understanding of cancer biology. Moreover, we delve into the role of biomedical imaging in monitoring disease progression and treatment response in these models. Biomedical imaging techniques offer a non-invasive approach to monitor tumor growth and therapy response. Modalities such as MRI/MRS/MRSI, PET, CT, and optical imaging provide valuable insights into tumor biology and the microenvironment. The integration of molecular imaging probes may enhance sensitivity and specificity in cancer detection. Within this special issue, we discuss the latest advancements in biomedical imaging for preclinical cancer research and their utility in guiding personalized treatment strategies.