Flow Cytometric Analysis of Inflammatory Cells in Experimental Acute Pancreatitis

Background: Accumulating evidence indicates that inflammatory cells migrate into the pancreas tissue and play an important role in the pathogenesis of acute pancreatitis (AP). The aim of this study was to establish a flow cytometric method to enumerate these infiltrating cells in the pancreas of an experimental AP. Materials and Methods: Twelve hours after inducing of AP, mice pancreatic tissues were cut into small fragments and single cells were prepared by mechanical dissociation. The isolated cells were stained with either anti-mouse CD45-PerCP or isotype antibody and analyzed by flow cytometry. Using side scatter (SSC)/CD45 gating we were able to identify inflammatory cells from non-inflammatory cells. Results: The mean percentage of leukocytes was 5.9±1.6 in the control group whereas, it was 26.7±8.1 in the AP. Moreover, we found that the percentage of lymphocytes, monocytes and granulocytes were 1.1±0.2, 0.9±.04 and 2.9±1.8 of total pancreatic cells, respectively, in the control mice. In contrast to lymphocytes, the percentage of monocytes and granulocytes were significantly increased in the AP group and it was 3±1.3 and 18.2±3.2 for monocytes and granulocytes, respectively. Conclusion: Quantitative flow cytometric analysis is feasible and provides a reliable and rapid assay to determine the number and percentage of inflammatory cells in experimental AP. K e y w o r ds: Acute pancreatitis, flow cytometry, inflammatory cells