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Differentiation of adipose‐derived stem cells into urothelial and smooth muscle cell lines within the structure of collagen/hyaluronan scaffold

Many urethral pathologies require surgical interventions. Although the commonly used surgical techniques are still being developed, complications such as the urethral strictures often occur and repeated surgery is needed. Tissue engineering of the urethral tissue, using autologous stem cells and appropriate scaffolds, represents promising technique to overcome these complications. In our experiment, we focused on the differentiation of adipose tissue‐derived stem cells seeded on the collagen/hyaluronan scaffolds into urothelial and smooth muscle cells. Differentiation culture media containing keratinocyte serum‐free medium with 2 % fetal bovine serum (FBS) and 30 ng/mL EGF, and D‐MEM with 10 % FBS, 2.5 ng/mL TGF‐β and 5 ng/mL PDGF were used to induce urothelial and smooth muscle differentiation, respectively. After two weeks of cultivation, cells were analyzed by scanning electron microscope (JEOL 7500F) for morphological features. Expression of the uroplakin 2 (UPK2) gene, keratin 2 gene (KRT2), ACTA 2 gene, and calponin 1 gene (CNN1) was tested in order to determine successful differentiation by quantitative Real‐Time PCR. Obtained results showed attachment of the cells on scaffold surface. The increased levels of UPK2 and KRT2 confirmed urothelial differentiation and expression of ACTA 2 and CNN1 confirmed smooth muscle differentiation. On the basis of obtained results, it can be emphasized that collagen/ hyaluronan scaffolds colonized by adipose tissue‐derived stem cells after performing further tests dealing with biosafety may have promising potential for tissue engineering of the urethra.