Figure 2
From: Organotypic pancreatoids with native mesenchyme develop Insulin producing endocrine cells
Organotypic pancreatoids develop Insulin producing beta cells. (a) Time course of organotypic pancreatoid development from Insulin1- EGFP mice. (b) Whole mount three-dimensional reconstruction of organotypic pancreatoid from a wildtype mouse at day 7 stained by immunofluorescence, with the epithelium marked by Pdx1 (in green) and beta-like cells marked by Ins (in red). Scale bar = 50 um. (c) Whole mount three-dimensional reconstruction of organotypic pancreatoid from a wildtype mouse at day 7 stained by immunofluorescence, with the acinar-like cells marked by Amy (in red), beta-like cells marked by Ins (in green), and endocrine progenitors marked by Ghrl (in white). Nuclei are marked by dapi in blue. Scale bar = 50 um. (d) Quantification of acinar-like and beta-like cell proportions. N = 8. Error bars are SEM. (e) Whole mount three-dimensional reconstruction of organotypic pancreatoid from a wildtype mouse at day 7 stained by immunofluorescence, with the beta-like cells marked by Ins (in green) and endocrine progenitors marked by Isl1 (in red). Nuclei are marked by dapi in blue. Scale bar = 50 um. (f) Fold change relative to 2.8 mM glucose showing c-peptide secretion during GSIS normalized to Ins+ cell number. N = 3. Error bars are SEM. (g) Absolute values of c-peptide secretion during GSIS normalized to Ins+ cell number. N = 3. Error bars are SEM. (h) Quantitative PCR to determine gene expression of key pancreatic markers, including Pdx1 (epithelium), Mist1, Prss1, and Hes1 (exocrine), and Pax6, Hnf6, Isl1, Nkx6-1, Ins1, and Ins2 (endocrine). Y-axis scale is log10. Expression is normalized to Gapdh. N = 4 for pancreatoids, N = 3 for murine tissue. Error bars are SEM.
