Generation of pancreatic endoderm and endocrine progenitor-like cells from iENDO.

<p>A) Protocol time-line for pancreatic endocrine differentiation from iENDO cells. B) Morphology of 3D aggregate at day 12 and day 30 of differentiation, 10x magnification (scale bar 50μm). C) Live/Dead staining of 3D aggregates at day 12 of differentiation (Live-green, Red-dead) (scale bar 50μm). D) H&E staining of day 12 iENDO pancreatic organoids. E)Relative gene expression (to <i>PPIG</i>, log scale) analysis represented in a heat-map of pancreatic endocrine marker 14TFs iENDO day 0, day 12 iENDO pancreatic organoids, day 30 3D iENDO pancreatic organoids and day 30 hESC pancreatic organoids compared with primary human Islets. All data represent mean of three independent experiments. F) Immunostaining for PDX1 (20X) in 293x transduced PDX1 (top) and day 12 iENDO pancreatic organoids (bottom) (scale bar 100μm, representative of N = 3) with respective isotype controls. F) Immunostaining for NGN3 (20X) in day 12 iENDO pancreatic organoids with respective isotype controls (scale bar 50μm, representative of N = 3). *p<0.05, **p<0.01 and ***p<0.001 determined by unpaired 2-tailed Student’s t-test.