Objective To construct the orthotopic transplantation tumor model with pancreatic cancer derived from transgenic LSL-Kras^G12D/+, LSL-Trp53^{R172 H/ +}, Pdx1-Cre(KPC) mice. To provide a stable and reliable preclinical animal model for studying the development mechanism and treatment strategy of pancreatic cancer. Methods The tumor tissue derived KPC transgenic mice with spontaneous pancreatic cancer was transplanted into C57BL/6J mouse pancreas. Ultrasound detection was used to monitor the growth of tumor. H E staining and immunofluorescence staining were used to evaluate the pathological characteristics of this model. Results The tumor derived from KPC mice could grow steadily on the pancreas of C57BL/6J mice. Tumor proliferation index Ki67, matrix fibrosis marker αSMA, immune cell markers CD45 and CD206 were all stably expressed in the tumor. The model stably retains the pathological features of primary pancreatic cancer. Widespread tumor metastases were development in this model, which was similar to those observed in patients with pancreatic cancer. Conclusion The orthotopic transplantation model derived from transgenic mouse with spontaneous pancreatic cancer was established successfully. The model can simulate the stromal environment and immune cell infiltration of pancreatic cancer, and retains strong stability and uniformity with original tumor. It can be used as an effective preclinical model to study pancreatic cancer progression and treatment strategies.