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Technically, pancreatic juice can be collected intraoperatively or with minimally invasive procedures, such as endoscopic ultrasound, retrograde cholangio-pancreatography, or by endoscopic collection of duodenal juice secretion 20. Several studies have explored the potential of pancreatic juice to identify novel biomarkers of disease using various approaches, including cytologic analysis 13, proteomic analysis performed by mass-spectrometry 14, 15, assessment of genetic and epigenetic markers such as K-ras and p53 mutations 16, 17, alterations in DNA methylation 18, and miRNAs 19. Due to its proximity to the pancreas, it could be strongly affected by microenvironmental perturbations induced by the tumor mass ( Figure 1), and therefore more informative than blood or urine, or tissue-based profiling. Pancreatic juice is a protein-rich fluid, which accumulates the secretome of pancreatic ductal cells and flows to the main pancreatic duct, and then to the main common bile duct. We recently demonstrated that pancreatic juice, a relatively unexplored body fluid, can be used to identify metabolic signatures of patients with distinct clinical profiles 12. Many studies have exploited comprehensive “omics” approaches, such as genomic, proteomic and metabolomic techniques, to identify candidate molecules or signatures that could discriminate between PDAC and other benign pancreatic afflictions. Promising results in biomarker discovery have been achieved using easily accessible body fluids, such as blood 5, 6, 7, urine 8, saliva 9 and pancreatic juice 10, 11, 12. The identification of variables associated to specific pancreatic pathologies could facilitate the surgical decision-making process and improve patient profiling. Furthermore, the overlap of symptoms between PDAC and other benign pancreatic pathologies can hamper the achievement of a prompt and reliable diagnosis with the current diagnostic strategies. The lack of specific symptoms until an advanced stage does not allow for an early diagnosis, contributing to the deadliness of the disease. Currently, surgical resection is still the only curative option for PDAC, yet there is a high frequency of early relapses and postsurgical complications. It is well-known for its immunosuppressive microenvironment and for its unresponsiveness to immunotherapy protocols 4. Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive tumors, and soon to become the second leading cause of death 1, 2, 3. We describe here two simple and technically undemanding methods for its isolation: tissue centrifugation and tissue elution. Since TIF is not readily accessible, various techniques have been proposed for its isolation. Similarly to pancreatic juice, for its property to collect and concentrate molecules that are found diluted in plasma, TIF can be exploited as an indicator of microenvironmental alterations and as a valuable source of disease-associated biomarkers. Tumor interstitial fluid (TIF) is the extracellular fluid, outside blood and plasma, which bathes tumor and stromal cells. Unfortunately, translating pancreatic juice collection to murine models of PDAC, to perform mechanistic studies, is technically very challenging.
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Here we describe in detail the intraoperative collection procedure. Pancreatic juice is a relatively unexplored body fluid, which, due to its close proximity to the tumor site, reflects changes in the surrounding tissue. There is an urgent need of variables associated to specific pancreatic pathologies to help preoperative differential diagnosis and patient profiling. Pancreatic adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death, and soon to become the second.