Tendon tissue engineering based on stem cell differentiation has attracted a great deal of attention in recent years. Previous studies have examined the effect of cell-imprinted polydimethylsiloxane (PDMS) substrate on induction differentiation in stem cells. In this study, we used tenocyte morphology as a positive mold to create a tenocyte-imprinted substrate on PDMS. The morphology and topography of this tenocyte replica on PDMS was evaluated with scanning electron microscopy and atomic force microscopy. Then the tenogenic differentiation induction capacity of tenocytes replica in ADSCs was investigated and compared with other groups including tissue replica (Which was produced similar to the tenocyte replica and was evaluated by SEM), decellularized tendon, and BMP-12 as other potentially inducers. This comparison gives us an estimate of the ability of tenocyte-imprinted PDMS (which called cell replica in the present study) to induce differentiation compared to other inducers. For this reason ADSCs were divided into 5 groups including control, cell replica, tissue replica, decellularized tendon and BMP-12. ADSCs were seeded on each group seperately and investigated by real-time RT-PCR after 7 and 14 days. Our results showed that in spite of the higher effect of growth factor on tenogenic differentiation, cell replica can also induce tenocyte marker expression (Scleraxis and Tenomodulin) in ADSCs. Moreover, tenogenic differentiation induction capacity of cell replica was greater than tissue replica. Immunocytochemistry analysis revealed that ADSCs seeding on cell replica for 14 days led to Scleraxis and Tenomodulin expression at the protein level. Also, immunohistochemistry indicated that contrary to the promising results in vitro, there was little difference between ADSCs cultured on tenocyte-imprinted PDMS and untreated ADSCs. The results of such studies could lead to the production of inexpensive cell culture plates or biomaterials that can induce differentiation in stem cells without growth factors or other supplements.