Figure 2. Organoids express markers associated with pancreatic progenitor cells.
(a) Expression of pancreas exocrine specific marker genes in 3D organoids and human fetal endodermal tissues. (b) Expression of markers for liver (ALB, CREBPA), stomach (SOX2) and duodenum (CDX2) in 3D organoids, human fetal pancreas and positive control (fetal liver or fetal stomach or fetal duodenum). (c) A schematic summarizing expression patterns of transcription factors during human embryonic pancreas development. Expressed genes are in red; repressed genes are in grey, adapted from Jennings et al10. (d) Expression of pancreas markers in 3D organoids, MCF-10A mammary epithelial cells used as non-specific control. (e) Expression of NKX2.2, a pancreatic endocrine marker gene. (f) Expression of markers in 3D organoids as detected by immunofluorescence. Images show representative fields and inserts show one organoid at a higher magnification. Scale bars, 50 µm. Graph summarizes results from three independent sets of experiments with over 100 organoids counted for each experiment. An organoid was designated as marker positive when more than 50% cells were positive for expression. (g) Expression of markers associated with differentiated ductal, acinar or islet cells. CA2, carbonic anhydrase II; CFTR, cystic fibrosis transmembrane conductance regulator; CEL, carboxyl ester lipase; PNLIP, pancreatic lipase; SPINK1, serine peptidase inhibitor 1; INS, insulin; GCG, glucagon. For all qPCR experiments data represent mean +/− S.E.M. P value (t-test, two tailed): N.S. - not significant; *P = 0.01 – 0.05; **P= 0.001 – 0.01; ***P = < 0.001 (N = 3 biological repeats).