Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells, thereby providing an experimental basis for cartilage repair. Methods Bone marrow mesenchymal stem cells (BMSCs) were cultured using the Percoll density gradient separation method, followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints, subjected to physical crushing, repeated freeze-thaw cycles, and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of decellularized scaffolds at different concentrations, three groups were designed with concentrations of 100% (Group A), 50% (Group B), and 30% (Group C), with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto the optimally concentrated scaffolds and cultured for one week to observe cell growth. Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45; osteogenic induction stained with alizarin red showed red calcific nodules; chondrogenic induction stained with alcian blue showed blue cartilaginous nodules; no apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin, eosin, and toluidine blue.There is a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds（P<0.05).The content of glycosaminoglycans is slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size, water absorption, porosity, tensile strength, and Young's modulus(P<0.05).After co-cultivation of stem cells with the scaffolds, cell adhesion was good. Conclusion Percoll density gradient separation can obtain high-purity rabbit BMSCs; the mixed decellularization method is more thorough. Group C (30%) scaffolds are the most suitable for constructing tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group C scaffolds.