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One major unresolved question in the field of pancreas biology is whether ductal cells have the ability to generate insulin-producing β-cells. Conclusive examination of this question has been limited by the lack of appropriate tools to efficiently and specifically label ductal cells in vivo. We generated Sox9CreER^T2 mice, which, during adulthood, allow for labeling of an average of 70% of pancreatic ductal cells, including terminal duct/centroacinar cells. Fate-mapping studies of the Sox9⁺ domain revealed endocrine and acinar cell neogenesis from Sox9⁺ cells throughout embryogenesis. Very small numbers of non-β endocrine cells continue to arise from Sox9⁺ cells in early postnatal life, but no endocrine or acinar cell neogenesis from Sox9⁺ cells occurs during adulthood. In the adult pancreas, pancreatic injury by partial duct ligation (PDL) has been suggested to induce β-cell regeneration from a transient Ngn3 …