Protocol to purify and culture human pancreatic cancer cells from patient-derived xenografts

Pancreatic ductal adenocarcinoma (PDAC) exhibits extensive inter- and intratumoral heterogeneity, along with a significant stromal component. This protocol outlines steps to generate patient-derived xenografts (PDXs), isolate and enrich human PDAC epithelial cells, and verify their identity using droplet digital PCR (ddPCR) and human-specific markers. It provides a robust approach for culturing cancer epithelial cells to develop a human PDAC model system. For complete details on the use and execution of this protocol, please refer to Grygoryev et al.1.