Tissue engineering technology offers an appealing approach for tissue reconstruction of the urothelium. Adipose‑derived mesenchymal stem cells (ADSCs) represent an abundant source for tissue engineering applications. However, ASCs primarily possess mesoderm lineage differentiation potential. It is difficult to induce differentiation of ASCs towards urothelial cells that are derived from the endoderm, although a recent findings have reported that a conditioned medium may drive ADSCs towards differentiation into the urothelium phenotype. In the present study, human ADSCs were isolated from abdominal adipose tissues and incubated in this conditioned medium for indicated time periods. Western blotting showed that protein expression levels of urothelial specific marks, including CK7, CK20 and UPIII, were increased after seven days' incubation, but immunofluorescence microscopy determined that cells with CK7 and UPIII staining were scarce, which suggested a low‑efficiency for the differentiation. Prolonging the incubation time did not further increase CK20 and UPIII expression. Furthermore, miR‑33a expression was increased with ADSC differentiation. Using synthetic miRNAs to mimic or inhibit the action of miR‑33a revealed that miR‑33a hinders the differentiation of ADSCs towards urothelial cells. Furthermore, luciferase reporter assay confirmed that β‑catenin and transforming growth factor‑β receptor (TGFR) are targets of miR‑33a. Inhibition of miR‑33a expression increased β‑catenin and TGFR expression and improved the efficiency of ADSCs towards differentiation into the urothelium phenotype. The present novel finding suggests that miR‑33 may be an important target in tissue engineering and regenerative medicine for urothelium repair.