Abstract：[Objective] To explore the repair effect of rabbit adipose-derived mesenchymal stem cell sheet on rabbit articular cartilage injury. [Methods] Rabbit adipose-derived mesenchymal stem cells were isolated and cultured in vitro, and the cells were identified by multi-line differentiation. The rabbit adipose-derived mesenchymal stem cell sheet was constructed by chemical method (by adding 50μg/ml ascorbic acid). After the cell sheet was successfully constructed, the following tests were carried: the structure of the cell sheet was observed under an inverted phase contrast microscope, and the gross observation and scanning electron microscope observation were also used. The rabbit knee osteochondral defect model (3.5mm in diameter and 3mm in depth) was prepared, and then the experiment was divided into two groups: control group and cell sheet group. Three months after the operation, specimens were taken for testing to observe the repair of cartilage. [Results] Primary rabbit adipose-derived mesenchymal stem cells grew adherently in a short spindle shape and triangle shape. After subculture, the cells gradually grew in a vortex shape. The third-generation rabbit adipose-derived mesenchymal stem cells had multi-lineage differentiation ability. The rabbit adipose-derived mesenchymal stem cells sheet had a multi-layer structure, and the cells secreted a large amount of extracellular matrix, and the cells were embedded in it. Three months after the operation, specimens were taken and correlative detections showed that a large amount of new cartilage tissue was regenerated in the osteochondral defect in the cell sheet group, and the shape and color of the new cartilage tissue were close to normal cartilage; In the control group, almost no new cartilage was formed. O’Driscoll score results also showed that the score of the cell sheet group was significantly higher than that of the control group, and the difference was statistically significant (P<0.05). [Conclusion] Rabbit adipose-derived mesenchymal stem cell sheet could promote the regeneration of rabbit articular cartilage and provided a new idea for the repair of clinical cartilage damage.