Fig. 2
Ectopic implantation of Cartilage progenitors recapitulates endochondral ossification. a Schematic of cell harvesting, characterization by cell surface markers, seeding in scaffolds, rhBMP-2 addition, and subcutaneous implantation. b–e Sample characterization at 2-4-8 weeks after implantation (n = 2–8). b Quantification of ossicle volume by computed tomography (CT). c Computed tomography images (CT, yellow circles indicate implant location) followed by gross morphology of dissected samples and details of conventional and tissue-specific histology staining (Red color at Safranin staining, and Blue color at Alcian Blue (A. Blue) staining, indicate cartilage extracellular matrix proteins; H&E, Hematoxylin and Eosin) and VEGF immunostaining. d Immunostaining to locate osteoclasts (VPP3), Vasculature (ENDOMUCIN) and implanted cells (TOMATO or YFP as indicated by white arrows). e Quantification of implanted cells within the newly formed cell phenotypes in 4-week samples (From tomato or YFP immunostaining. “Sox9-Tomato”, quantification from Tomato immunostaining at scaffolds implanted with tamoxifen-treated cells obtained from tomato reporter mice; “Col2A1- YFP”, quantification from YFP immunostaining at scaffolds implanted with cells extracted from Col2A1- YFP reporter mice)
