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Background: Sex hormones play important role in proliferation, differentiation, maturation and the scheduled death of chondrocytes. Although, some studies report the regulatory role of estrogen in the development and progression of cartilage, some of the mechanisms still remain unclear, including the role of estrogen in the expression of cartilage-specific genes in chondrogenesis process. We studied this role in the present study. Methods: We used adipose-derived stem cells, which were previously differentiated into cartilage tissue in pellet culture system in the control (without estrogen in culture medium) and experimental (with estrogen in culture medium) groups. Production of chondrogenesis markers, type II collagen and aggrecan were evaluated in experimental and control groups via immunohistochemical (IHC) technique. Then, the results were evaluated with Image-J software and statistical analysis were done. Findings: Estrogen led to decrease of type II collagen and increase of aggrecan production. Conclusion: This study showed that chondrogenesis could be affected by estrogen.