Abstract B124: Isolation and characterization of ascitic derived primary pancreatic cancer cell cultures from different patients as a platform for personalized medicine.
Authors: Golan Talia , Atias Dikla , Berger Raanan 
Published: June 2012

Discovery and development of effective cancer treatments in pancreatic cancer (PC) has been hampered by the lack of reliable predictive preclinical models to assess the therapeutic efficacy of candidate agents, mostly due to the genomic heterogeneity in these patients. Therefore alternative models that recapitulate this complex disease need to be developed. Approximately a third of patients with PC have a tendency to produce ascites in advanced disease. The ascitic fluid contains viable tumor cells, and therefore we decided to develop an investigational model based on primary tumor cells obtained from patients' ascites. Ascitic fluid was obtained from 20 patients with PC undergoing palliative paracentesis, in 4 patients ascites was obtained several times during the course of their disease (totaling 44 ascitic fluids obtained). Ascitic-derived primary pancreatic cancer cells were isolated and cultures were grown for each individual patient. We identified the PC cells and examined the cells' morphological characteristics, phenotypes, invasive properties and the effect of various therapeutic agents. We successfully established ascitic-derived primary cell cultures in 84% (37/44). Positive staining by IHC of epithelial marker (CK-7) and negative staining for CK-5/6 was demonstrated in the majority of cells in the culture. Cytokeratin staining by FACS was observed in 90–95% of the isolated cells supporting the presence of homogeneous epithelial PC enriched cell culture. We observed a wide range in doubling time and migration properties among the patients' ascites derived cell cultures and even among cultures from the same patient along the course of his disease. A correlation between high migratory potential and long doubling time was observed. The cells were treated with different chemotherapies and biological agents. The diverse nature of each individual patient's primary pancreatic cancer cell cultures were further demonstrated by varying responses to the therapeutic agents examined. Morphological changes were also observed at different stages of disease, most cells initially displayed a characteristic epithelial morphology, but with more advanced disease a mixed morphological appearance ‘epithelial-mesenchymal’ was seen. The expression of epithelial-mesenchymal-transition (EMT) markers was examined by real-time PCR. Up regulation of Zeb-1, Zeb-2, Twist, Slug and Dab2 were observed in the cells displaying ‘epithelial-mesenchymal’ like appearance. We have developed a unique primary ascitic-derived pancreatic cancer cell culture model that enables a diverse range of patients-related cell cultures. These cell lines has the potential to serve as a relevant model to study signaling pathways in PC progression. Additionally, this model could be used to assess the sensitivity to therapeutic agents in a short time frame, therefore supporting personalized treatment decisions.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr B124.