Effect of Reg Protein on Rat Pancreatic Ductal Cells

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Abstract

Summary

The pancreatic regenerating gene (reg I) is expressed in the exocrine pancreas and is involved in islet regeneration. reg I protein has been shown to be mitogenic to p- and ductal cell lines, but not mature islets. In this study, we tested the effect of two isolates of reg I on primary cultures of ductal cells. Rat pancreatic ductal cells were isolated by collagenase digestion and isolated colonies were maintained in culture. The cells were proven to be ductal in origin by their morphology and by immunofluorescent staining with epithelial markers. reg I was isolated from human pancreatic extracts or from the rat acinar cell line AR42J by sequential ammonium sulfate precipitation and acid precipitation. Cells were cultured with doses of reg I for 72 h, pulsed with 10 μM bromodeoxyuridine (BrdU) for 2 h. After fixation, nuclei were double-stained with propidium iodide and BrdU monoclonal antibody. The percentages of nuclei positive for BrdU were calculated from at least five colonies per group. A 10-nM concentration of human reg I increased BrdU incorporation by 2.3-fold over controls, rat reg I increased it by 1.4-fold (p <0.05). When compared to their effects on the ductal cell line ARIP, both human and rat reg I were 100 times more potent on the primary cultures of ductal cells. We conclude that human and rat reg I proteins are mitogenic to primary cultures of ductal cells. Although principally a product of the acinar cell, reg I appears to be a stimulus of ductal cell growth and, in this fashion, may modulate the expansion of the pancreatic ductal population during islet regeneration.

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