Primary Culture of Pancreatic Acinar Cells in a 3-D Collagen Matrix
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Authors
Emeott, Kayla
Issue Date
2010
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
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.
Description
v, 21 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.