Introduction & Objectives: Porous scaffolds derived from native cartilage matrix along with autologous cells could be an effective tool for cartilage tissue engineering (CTE). Recently, it was shown that scaffolds based on cartilage extra cellular matrix (ECM) can induce chondrogenesis of human adipose-derived mesenchymal stromal cells (ASCs) without using exogenous growth factors. However, lack of mechanical properties, rapid biodegradation, and contraction of these scaffolds in culture limit further applications. The present study investigated the fabrication of novel scaffolds based on devitalized costal cartilage matrix (DCM) and poly vinyl alcohol (PVA), using genipin as a natural crosslinker. The scaffolds evaluated in vivo subcutaneously in Vistar rats.Methods and Materials: For this purpose, PVA was modified to expose amine groups (PVA-A), which crosslinked with DCM powder via the lowest genipin percentage of 0.04 % (wt/wt). The crosslinked scaffolds were characterized by different techniques including porosity percentage, pore size, mechanical properties, crosslinking density, and swelling. ASCs were seeded on the scaffolds using fibrin hydrogel. The scaffolds implanted in dorsum of Vistar rats for 28 days.Results: After 28 days the animals sacrificed. Gene expression measurements, biochemical assays and histological staining confirmed that ASC-seeded constructs cultured in the chondrogenic medium can express cartilage-specific genes and synthesize cartilage-related macromolecules. In the presence of TGF-β3 the constructs exhibited significant expression of these markers compared to the control medium.Conclusion: These findings suggest that [genipin-crosslinked DCM-PVA-A/ fibrin] can be considered as an appealing hybrid scaffold for CTE applications. According to the results. the ex vivo strategy can consider as versatile method for cartilage engineering.