The primary ADSCs of SD rabbits were extracted for three-dimensional differentiation and flow cytometry was used to detect the cell dryness.The ear cartilage of 3-month-old New Zealand white rabbits was collected and treated with grinding and soaking method to prepare ACM. The experiment was divided into ACM and ACM+ADSCs co-culture group.The ultrastructures of ACM and the ADSCs in ACM were observed under scanning electron microscope，and the growth status of ADSCs in ADSCs+ACM co-culture group was by Calcein-AM/PI fluorescence staining method. The ADSCs were divided into control group and experimental group（ADSCs and ACM co-culture）.The CCK-8 method was used to detect the proliferation activities of cells in two groups. Real-time fluorescence quantitative PCR （RT-qPCR） was used to detect expression levels of type Ⅱ collagen （COL-Ⅱ），type Ⅹ collagen （COL-Ⅹ）， SRY-related high mobility group-box 9 （SOX-9），vascular endothelial growth factor （VEGF）， alkaline phosphatase （ALP） and Runt-related transcription factor 2 （RUNX-2） mRNA in the ADSCs in two groups at 7 and 14 d of chondrogenic induction.

The rabbit ACM was successfully prepared. The extracted ADSCs could differentiate in three ways and the specific surface markers of stem cells were expressed in the ADSCs. The ADSCs could adhere to ACM. Compared with control group， the proliferation activity of the cells in experimental group was increased （P<0.05）.The RT-qPCR results showed that compared with control group， the expression levels of VEGF，ALP and RUNX-2 mRNA in the ADSCs in experimental group were increased at 7 d of chondrogentic induction （P<0.05）， and the expression levels of COL-2， SOX-9， and COL-Ⅱ mRNA had no significant differences（P>0.05）. At 14 d of chondrogentic induction， compared with control group， the expression levels of VEGF， ALP， and RUNX-2 mRNA in the ADSCs in experimental group were increased （P<0.05），and the expression level of COL-Ⅹ mRNA was decreased （P<0.05）. Compared with 7 d of chodrogenic induction， the expression levels of VEGF， ALP， and RUNX-2 mRNA in the ADSCs in control group at 14 d of chodrogenic induction were decreased （P<0.05）， and the expression levels of COL-Ⅱ and COL-Ⅹ mRNA were increased （P<0.05）. Compared with 7 d of chondrogenic induction， the expression levels of COL-Ⅱ and COL-Ⅹ mRNA in the ADSCs in experimental group were increased at 14 d of chondrogenic induction （P<0.05）， while the expression levels of VEGF， ALP， and RUNX-2 mRNA were decreased（P<0.05）.

The rabbit ACM lack immunogenicity and have good biocompatibility and chondrogenic induction ability， which is beneficial to the occurrence of endochondral osteogenesis.