Objective To construct a patient-derived pancreatic tumor organoid ( PDO) and evaluate its effectiveness. Methods We collected fresh surgical specimens from pancreatic cancer patients for PDO culture and compared the pathological and genetic characteristics of the PDO model with those of primary tumors. The PDO model was used to evaluate the efficacy of clinical chemotherapy drugs, and the effectiveness of the model was assessed. Results A PDO model of pancreatic cancer was successfully established. Histomorphological analysis indicated that the PDO model maintained the basic pathological characteristics of the primary tumor. Whole-exon sequencing showed that both the organoids and original tumor tissue remained consistent in their gene mutation type and characteristics. Drug screening tests revealed that the PDO model had good sensitivity to gemcitabine and irinotecan. Conclusions A pancreatic cancer PDO was successfully constructed that reflected the histological and genetic characteristics of the original tumor. The model was shown to be effective for drug sensitivity experiments in vitro and is expected to have implications for precision medicine assays.