Abstract only Many lizards are capable of spontaneous tail regeneration following tail loss. Although the regenerate tail superficially resembles the original, it is not identical. One of the main differences is that a cartilaginous cone replaces the original bony vertebrae of the axial skeleton. Here we characterize spontaneous chondrogenesis during tail regeneration in the leopard gecko. Following tail loss, the first sign of regenerative outgrowth is the formation of a blastema, an aggregation of proliferating mesenchymal‐like cells localized at the site of tail loss. Cartilage regeneration begins within the blastema 8–14 days post‐injury. Presumptive cartilage first appears as a glycosaminoglycan (GAG) and type II collagen (Col2) positive condensation that surrounds the newly regenerating spinal cord. With continued regenerative outgrowth, this condensation forms a hollow cone‐like structure. Presumptive cartilage cells express phosphorylated Smad2 (pSmad2), indicating activation of the canonical transforming growth factor beta (TGFβ) signalling pathway. Curiously, early pSmad2 expression is not matched by TGFβ1 or TGFβ3 expression, indicating that a different member of the TFGβ pathway is involved in early chondrogenesis. Furthermore, the transcription factor Sox9, often described as the master regulator of chondrogenesis, is not expressed until several days after the first appearance of Col2 and GAG in the newly forming tail. Chondrogenesis is complete within ~30 days, yielding a distinctly cell‐rich/matrix‐poor structure. Our findings indicate that regeneration‐mediated chondrogenesis in the lizard tail is not a simple recapitulation of embryonic cartilage development. Support or Funding Information Natural Sciences and Engineering Research Council (NSERC) Discovery Grant 400358