ABSTRACT
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