Pancreatic ductal adenocarcinoma (PDA) organoids are 3D cultured from patient-derived stem cells or progenitor cells in vitro. PDA organoids have a variety of cell types, can realize structural self-organization through cell self-renewal, and are similar to the cells in the body of the original organ function in vivo biological bank. PDA organoids can be derived from surgical or biopsy tissue. The ability to build organoids from biopsy will facilitate the sampling of a larger population of PDA patients. Repeated sampling of patients can track the entire progression of the disease longitudinally. Compared with the traditional 2D cell culture and patient-derived xenotransplantation models, the three-dimensional culture of PDA organoids has the characteristics of short time and high success rate, and can be cryopreserved and maintain the stability of genetic traits. Organoids that can simulate diseases can be used as an alternative drug testing system. Using it for drug testing can not only better reflect the patient's response to drugs, but also can reduce the number of animal experiments. Moreover, when using organoids for testing, there is no need to understand the underlying molecular mechanism a priori, and chemical sensitivity testing can be performed directly, thereby shortening the testing time. In this paper, the advantages and disadvantages of different PDA organoids 3D culture methods and the verification methods for the stability and invasiveness of PDA organoids were reviewed. The mechanism of PDA organoids used for tumor chemotherapy drug sensitivity screening was discussed, and the application prospects and challenges of tumor biology in patient individualized treatment and precision medical treatment were discussed.