Primary Culture of Pancreatic Acinar Cells in a 3-D Collagen Matrix
Pancreatitis is a common, potentially life-threatening inflammatory disease of the pancreas. It is widely accepted that the premature activation of digestive enzymes within the pancreatic acinar cell, the main functional cell of the exocrine pancreas, is a critical initiating event in the development of this disease. For this reason, the acinar cell represents a key experimental model system to study disease mechanisms. Unfortunately, there has been little success maintaining these cells in culture for more than four days. The aim of this study was to establish a reliable technique for the long term culture of pancreatic acinar cells. We predicted that culturing the cells in a three dimensional collagen matrix would be ideal for maintaining cells since in the body, they are in direct contact with collagen in the extracellular matrix. Acinar cells were isolated from rat pancreata and cultured in a three dimensional collagen matrix in DMEM culture media. The cells were then evaluated for health over time by examining their polarity. Cell polarity was examined by immunofluorescently staining F-actin which is apically located in healthy cells. Cell health degraded after only 24 hours, however and it was concluded that this might be due to a flaw in the isolation technique and that the method should be revised to better the chances of successfully culturing cells. The method was optimized to reduce variability in cell size and density inter- and intra- experimentally by introducing cell size exclusion steps into the methods and identifying an ideal cell density for consistently obtaining assay results with smaller standard deviations. The data obtained for this optimization, however, was not normalized so conclusions about its success could not be made.
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