In our previous investigations, we have proved that the viscoelasticity of the Articular chondrocytes undergoes dramatic changes during their normal development, however, whether the temperature, one of the important factors in cell cultivation and cartilage tissue engineering, is related with the viscoelasticity of cell in not known. The goal of this study was to characterize the changes of viscoelastic properties of chondrocytes in different age group under different temperature. The micropipette aspiration combined with a standard linear viscoelastic solid model was used to quantify the effect of temperature changes on viscoelastic properties and deformation recovery of chondrocytes isolated from different age groups. Results have shown that temperature significantly influences the viscoelastic properties of chondrocytes isolated from different group and the deformation recovery of which have significantly changed with temperature changing. Moreover, we found that the cytoskeletal network were more sparsely, and the contents of the components were reduced in old cells determined by immunohistochemistry, LSCM, real time PCR and western blotting compared with the adult and the young cells. These findings provide substantial temperature-related information which may be relevant for chondrocyte-based cartilage tissue engineering.