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Breast cancer is the most prevalent malignant tumor in women and the second leading cause of cancer-associated mortality among female patients. Characterized by prominent subtype heterogeneity, breast cancer arises from stepwise accumulation of gene mutations in mammary epithelial cells, exhibiting complex genomic profiles and divergent biological phenotypes. At present, standard clinical therapeutic regimens are formulated mainly according to patients’ pathological features and the expression status of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). Such strategies fail to fully match the tumor characteristics of individual patients, imposing considerable restrictions on individualized precision therapy. Accordingly, systematic elucidation of normal mammary gland development, tumor initiation mechanisms and malignant progression is of great significance for developing novel personalized therapeutic approaches. Compared with conventional breast cancer cell line models, breast cancer organoids support long-term stable in vitro culture and can faithfully recapitulate the histopathological features, hormone receptor expression patterns, HER2 status, copy number variations and gene mutation landscapes of patients’ primary tumors, maximally preserving the tumor heterogeneity of breast cancer. These organoid models can be applied to in vitro evaluation of drug efficacy; meanwhile, the establishment of in vivo xenograft models enables verification of the therapeutic effects of novel anti-tumor agents. Combined with the mammary imaging window technique, real-time dynamic monitoring of xenograft growth and invasive behaviors of tumor cells can be achieved. Most importantly, breast cancer organoids steadily retain the genetic background and histological characteristics of matched primary tumors, allowing accurate testing of individualized drug sensitivity. They serve as an excellent preclinical platform to facilitate mechanistic research on breast cancer and advance the clinical translation of precision oncology.