Abstract: Objective:To investigate the chondrogenic differentiation of umbilical cord derived mesenchymal stem cells isolated and cultured from cryopreserved human umbilical cord (hUC-MSCs) .Methods:hUC-MSCs isolated and cultured from cryopreserved human umbilical cord were divided into the induction group and control group, and were cultured in complete chondrogenic differentiation medium and conventional medium for 7 days, respectively. The expression of chondrogenic markers Col Ⅱ a1, Col X a1 and Aggrecan mRNA in the hUC-MSCs was examined by real-time fluorescence quantitative (qRT) -PCR, and the expression of collagen type Ⅱ protein in the hUC-MSCs was examined by immunoblotting analysis and immunofluorescence staining. The content of glycosaminoglycans (GAG) in the hUC-MSCs was examined by Alcian blue staining.Results:After 7 days of chondrogenic differentiation, the hUC-MSCs showed higher mRNA expression of Col Ⅱ a1, Col X a1 and Aggrecan, higher protein expression of collagen type Ⅱ, and higher content of GAG, compared with the control group (all P<0.05) . Conclusion:hUC-MSCs isolated and cultured from cryopreserved human umbilical cord can differentiate into chondrocytes after 7-day induction in chondrogenic differentiation medium. This can be useful for experimental research and clinical treatment on cartilage diseases.