Figure 4.

Endocrine Cell Markers in Pancreatic Organoids after 7 Days of In Vitro Differentiation

(A) Bright-field image of human pancreatic organoids initially expanded for 7 days followed by differentiation culture for 7 days. DAPI was used as nuclear counterstain. Scale bar, 100 μm.

(B) Confocal image of pancreatic organoids immunostained for Ki67 (green) and KRT19 (red). The majority of cells were KRT19⁺. No cells were Ki67⁺. DAPI was used as nuclear counterstain. Scale bar, 50 μm.

(C) Confocal image of pancreatic organoids immunostained for KRT19 (green) and INS (red). Few INS⁺ (0.52%) cells were present. DAPI was used as nuclear counterstain. Scale bar, 50 μm.

(D) Confocal image of pancreatic progenitor markers PDX1 (green) and SOX9 (red). DAPI was used as nuclear counterstain. Scale bar, 50 μm.

(E) Confocal image of pancreatic progenitor marker NEUROG3 (red). DAPI was used as nuclear counterstain. Scale bar, 50 μm.

(F) Confocal image of pancreatic progenitor marker NKX6.1 (green). DAPI was used as nuclear counterstain. Scale bar, 50 μm.

(G) Gene expression of several markers in organoids on day 7 of differentiation compared with organoids on day 7 of expansion. Gene expression in expansion organoids were set to 1 (dotted line). Mean ± SEM, n = 8 donors. ^∗p < 0.05 and ^∗∗p < 0.01.

(H) Percentage of cells immunostained for different markers on day 7 of expansion (white bars) compared with day 7 of differentiation (black bars) using confocal images. More than 10 organoids per donor were assessed. Mean ± SEM, n = 4–6 donors. ^∗p < 0.05 and ^∗∗p < 0.01.

See also Figure S4.