INTRODUCTION: Bladder urothelial carcinoma is highly heterogeneous and is prone to recurrence. There are many types of drugs for intravesical instillation, and there is an urgent need for reliable tumor models for individualized treatment strategies. Patient-derived organoids (PDOs) are efficient and accurate neotype tumor models that can compensate for the deficiencies of traditional tumor models. METHODS: From September 2023 to February 2024, bladder cancer tissues were collected from surgical resections in the Department of Urology, Beijing Tiantan Hospital, Capital Medical University. We explored suitable culture systems and cultured primary tumor cells in Matrigel matrix gels. Hematoxylin and eosin (HE) staining and immunohistochemistry were performed for comparison with the original tumor tissue at the histological level for validation. Drug sensitivity tests were also conducted. RESULTS: In this study, a culture system for PDOs was successfully established. HE staining and immunohistochemistry were used to morphologically verify the organ model. Drug sensitivity tests have clarified the value of PDOs as preclinical models of precision medicine. CONCLUSION: A bladder tumor PDO culture platform was initially established, which laid a foundation for the subsequent construction of the organoid bank, the study of bladder tumor progression mechanism, and research in precision medicine.