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Fig. 3. Ex vivo differentiation of pancreatic precursors. Double immunostaining for CPA (green) and E-cad (red) (A,C) or for insulin (green) and glucagon (red) (B,D) in pancreatic explants on day 0 (A,B) and on day 7 (C,D) in culture. Nuclei are stained blue with Hoechst in B and D. (A) On day 0, only a few of the epithelial cells stained for E-cad express CPA and most of these are located at the periphery (yellow). These CPA positive cells are not polarized and are not organized into acini. (B) On day 0, there are small clusters of glucagon-expressing cells and very few insulin-expressing cells. (C) On day 7, epithelial cells strongly expressing CPA form acini with a characteristic polarization of the cells and a central lumen (inset). Ductal structures that are negative for CPA (arrowheads in C) are also observed. (D) On day 7, insulin-expressing cells, which have appeared during the culture, and glucagon-expressing cells are organized in large endocrine clusters. Scale bars, 100 µm (A-C); 50 µm (B).
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