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Summary: This protocol is a procedure for generating orthotopic isografts using mouse pancreatic cancer organoids. These isografts can be used to track the evolution of pancreatic ductal adenocarcinoma (PDA) from a preinvasive lesion to a metastatic disease and therefore represent a suitable model for identification of determinants of PDA progression.For complete details on the use and execution of this protocol, please refer to Boj et al. (2015) and Filippini et al. (2019).