AIM: A broad-range proteomic approach was applied to investigate the complexity of the mechanisms involved in pancreatic regeneration for identification of new treatment targets and potential markers of pancreatic stem cells. METHODS: A regeneration pancreatic model was induced by partial (90%) pancreatectomy in rats. Changes in protein expression in rat regeneration pancreas at 3 d after partial pancreatectomy, as compared to sham surgery, were analyzed using two-dimensional gel electrophoresis (2-DE), mass spectrometry (MS), and mass fingerprinting. RESULT: Two-DE displayed 91 spots with at least 1.5-fold of differentially expression at the time point of 3 days after pancreatectomy and 53 differentially expressed proteins were identified by peptide mass fingerprinting(PMF). These included embryogenic and cell proliferation-related, lipid and energy metabolism-related, protein and amino acid metabolism-related proteins, signal transduction proteins. CONCLUSION: The proteome profiling technique provided a broad-based and effective approach for the rapid assimilation and identification of adaptive protein changes while pancreas regeneration induced by pancreatectomy. Our data highlight the globe proteome during the pancreatic proliferation and differentiation processes, which is very important and will lead to better understand regulation mechanism of the pancreatic regeneration, and to ultimately reach the target of discovering protein biomarkers of pancreatic stem cells.