Fig. 6: Recapitulation of disease phenotypes of OI.
From: Jawbone-like organoids generated from human pluripotent stem cells
a, Immunofluorescence staining for COLI and CHP in OI- and gene-corrected (rescued) OI (resOI)-iPSC-derived aggregates after 3-day osteogenic induction. Nuclei stained with DAPI (n = 6 from 3 biologically independent experiments). Scale bars, 25 μm. b, HE and Azan staining in serial sections of d38 OI- and resOI-jawbone-like organoids (n = 6 from 3 biologically independent experiments). Scale bars, 100 μm. c, Phalloidin staining in OI- and resOI-organoids. Images represent z-stack projection of 40 confocal optical sections (step size, 0.5 μm). Dashed boxes magnified on the right (n = 6 from 3 biologically independent experiments). Scale bars, 25 μm. d, 3D reconstructed micro-CT images of ectopic mineralized tissues in the renal regions of NSG mice at 4 weeks after OI- or resOI-organoid transplantation. Scale bars, 1.0 mm. e, HE and Azan staining of the transplanted tissues (n = 6 from 3 biologically independent experiments). Scale bars, 100 μm. f, Quantification of mineralized tissue volume and TMD in OI- and resOI-organoid transplanted groups. g, Quantification of osteocytes and Azan Red-stained areas. h, TUNEL staining of the transplanted tissues in OI- and resOI-organoid transplanted groups (n = 6). Dashed boxes magnified below. Scale bars, 50 μm. i, Immunofluorescence staining for human Vimentin in the transplanted tissues. F-actin stained with phalloidin. Images represent z-stack projection of 40 confocal optical sections (step size, 0.5 μm) (n = 6). Scale bars, 25 μm. j, SHG imaging for collagen fibres in the bone tissues. F-actin stained with phalloidin (red). Images represent z-stack projection of 23 confocal optical sections (step size, 1.0 μm) (n = 6). Scale bars, 25 μm. k, Quantification of TUNEL-positive apoptotic cells. l, Quantification analysis of osteocyte dendritic process number. m, Quantification analysis of SHG intensity normalized to resOI. In f, g and k–m, data are shown as mean ± s.d. (n = 6; except for m, n = 5). P < 0.05 by two-tailed unpaired Student’s t-test. In a–e and h–j, representative data are shown. All specimens stained after decalcification.
