Abstract
e14646 Background: In Pancreatic Cancer (PC) challenges in drug development include: obtaining metastatic cancer tissue for research, developing and validating biomarkers predicative for personalized therapeutic decisions. We have recently developed a novel therapeutic model for PC to address these challenges based on the isolation of viable PC cells derived from ascitic fluid. Methods: Ascitic fluid was obtained from PC patients undergoing palliative paracentesis. In several patients ascites was obtained several times during the course of their disease. Ascitic-derived primary PC cells were isolated, cultured and identified for each patient in vitro. Informed consent was obtained for all patients. Results: We successfully established ascitic-derived primary cell cultures in 87% (53/61) of the ascites obtained. Homogeneous epithelial PC enriched cell cultures were identified; CK 7/8 (epithelial marker) staining by FACS in 90-95% of the cells, IHC strong staining of CK 7 and negative staining for CK 5/6 in the majority of cells in the cultures. We observed a wide range in doubling time and migration properties among the patients' cell cultures. The diverse nature of each individual patient's cell cultures were further demonstrated by varying chemo-sensitivity to gemcitabine and additional therapeutic agents. Morphological changes were observed at different stages of disease, most cells initially displayed a characteristic epithelial morphology. With more advanced disease a mixed morphological appearance "epithelial-mesenchymal" was seen and up regulation of various epithelial-mesenchymal-transition markers were observed by real-time PCR. Conclusions: We have developed a unique primary ascitic-derived PC cell culture model. These cell lines have the potential to serve as a relevant model to study signaling pathways in PC progression, and to assess the sensitivity to therapeutic agents in a short time frame, therefore supporting personalized treatment decisions.
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