Figure 3. Differentiation of pancreatic progenitor organoids in vitro and in vivo.
(a) Schematic representation of the protocol for inducing progenitor organoids into ductal and acinar cells in culture. PTOM refers to Pancreatic Progenitor and Tumor organoid Media; PODM I and PODM II refer to Pancreatic Organoid Differentiation Media I and II. (b) QPCR analysis for ductal (CA2) and acinar (CPA1) markers. Immunofluorescence analysis of CA2 (green) and CPA1 (red) expression. (c) Top panel: H&E morphology of mammary ducts of control glands (left) and immunofluoresence for human leukocyte antigen (HLA), (right, HLA-Red). Bottom panel: organoid transplants stained with H&E (left) or for HLA (right). Transplants were identified in 20 out of 22 glands analyzed. (d) Comparative analysis of organization of H&E stained human fetal pancreas (top) with transplants (bottom) and immunofluorescence using ductal (KRT19, green), and acinar (CPA1, red) markers. (e) QPCR analysis showing expression of acinar (CPA1, CEL) and ductal (CA2, CFTR) markers relative to the progenitor organoids in transplants. (f) NKX6.1 expression in, progenitor organoids in culture (top) and ducts in transplants (bottom). DAPI, blue; NKX6.1, green; SOX9, red. (g) Transplants immunostained for ductal markers CA2 (red), KRT19 (cyan), primary cilia (acetylated tubulin, green) and DAPI (blue). (h) Transplants immunostained for ductal marker, primary cilia (acetylated tubulin, green) and acinar marker (CPA1, red) and DAPI (blue). All scale bars represent 50 µm. P = 0.01 – 0.05; **P= 0.001 – 0.01; ***P = < 0.001 (N = 3 biological repeats).