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Pancreatic islet transplantation is a viable treatment for type 1 diabetes, but is limited by human donor tissue availability. The combination of epidermal growth factor (EGF) and gastrin induces islet β-cell neogenesis from pancreatic exocrine duct cells in rodents. In this study we investigated whether EGF and gastrin could expand the β-cell mass in adult human isolated islets that contain duct as well as endocrine cells. Human islet cells were cultured for 4 wk in serum-free medium (control) or in medium with EGF (0.3 μg/ml), gastrin (1.0 μg/ml), or the combination of EGF and gastrin. β-Cell numbers were increased in cultures with EGF plus gastrin (+118%) and with EGF (+81%), but not in cultures with gastrin (−3%) or control medium (−62%). After withdrawal of EGF and gastrin and an additional 4 wk in control medium, β-cell numbers continued to increase only in cultures previously incubated with both EGF and …