Fig. 1: Micropatterned gut spheroid generator (μGSG) enhances the biogenesis of posterior foregut (PFG) spheroids from human pluripotent stem cells (hPSCs).
From: Mechanically enhanced biogenesis of gut spheroids with instability-driven morphomechanics
a Schematic showing endoderm patterning and region-specific organogenesis of the gut tube. PFG: posterior foregut. dAFG: dorsal anterior foregut. vAFG: ventral anterior foregut. MHG: mid-hind gut. b Schematic and representative phase contrast images of conventional monolayer-based induction of gut spheroids through an intermediate definitive endoderm (DE) stage. Scale bar: 100 μm. n = 3 independent experiments. c Schematic of the generation of gut spheroids in μGSG. d Induction of PFG spheroids. e Representative fluorescence image of micropattern (left) and phase contrast images of cell culture in μGSG on day 5 (middle) and day 7 (right). Scale bar: 500 μm. n = 3 independent experiments. f Representative time-lapse confocal micrographs showing PFG spheroid morphogenesis from days 6 to 7. (Inset) Schematic showing the optical section selected for viewing tissue fission. H1 H2B-mCherry cells and μGSG with a diameter of 200 μm were used. White arrowheads indicate newly formed spheroid. Yellow arrows indicate the tissue fission site. Scale bar: 200 μm. Similar results were seen in n = 3 independent experiments. g Representative phase contrast images showing PFG spheroids collected from monolayer-induction (left) and μGSG (right) on day 7, respectively. Scale bar: 500 μm. h Bar plot showing the number of PFG spheroids generated per 300,000 hPSCs input by monolayer-induction and μGSG, using three hPSC lines (iPS18, H1, and iPSB1), respectively. Data were plotted as mean ± s.d. n = 3 independent experiments. i Representative confocal micrographs showing immunostaining of indicated markers in μGSG-derived PFG spheroids. DAPI (blue) stains the nuclei. Scale bar: 50 μm. j Scatter plot showing the percentages of cells positive of indicated markers in PFG spheroids derived from monolayer-induction and μGSG, respectively. Data were plotted as mean ± s.d. n = 5 independent experiments. P-values were calculated using unpaired, two-sided Student’s t-test. Source data are provided as a Source Data file.
