Figure 4. Transplanted HLO-scaffolds consisted of mesenchymal cells and vasculature.

(A) Airway-like structures were surrounded by myofibroblasts, PDGFRα+ (green) and SMA+ (white) as well as smooth muscle, PDGFRα-/SMA+ (white only). Scale bars represent 50 µm in the lower mag image (left panel) and 25 µm in the lower mag image (right panel). (B) SafraninO staining showed clusters of cartilage in the 8 wk tHLO (right panel) but not in the scaffold grown in vitro for 8 weeks (left panel) Scale bar represents 100 µm. (C) Some βCAT+ (red) cells surrounding the airway-like structures expressed vasculature marker PECAM (white), but did not express human nuclear marker (HUNU, green) indicating that the vasculature is of host origin. Scale bars in A represent 50 µm lower mag image (left panel) 25 µm in bottom panel in the lower mag image (right panel). The low mag image scale bar represents 50 µm and high mag represents 10 µm. (D) 3D rendering of z-stack images on thick 12 wk tHLO sections (derived from H9 hESC) revealed the PECAM+ vascular network (white) around the SOX2+ airway epithelium (green). Scale bar represents 100 µm.

DOI: http://dx.doi.org/10.7554/eLife.19732.017

Figure 4—figure supplement 1. Cartilage observed in transplanted HLOs is of human origin.

The cartilage in the tHLO expressed SOX9 (green) and huMITO (white). Scale bar represents 50 µm in the low mag image and 10 µm in the high mag image. (B) 15d HLOs grown in vitro only possess clusters of SOX9+ cells in the mesenchyme, suggesting HLOs possess cartilage precursor cells. Scale bar represents 50 µm.

DOI: http://dx.doi.org/10.7554/eLife.19732.018