A Preliminary Study on the Promotion of Canine Adipose-Derived Stem Cell Differentiation by Perfusion-Decellularized Ureter Matrix

Abstract

This study was to assess the possibility of the perfusing decellularized ureters (DUs) promoting the differentiation of the canine adipose stem cell (cASCs). cASCs were isolated and cultured in different induction media to determine their multidirectional differentiation potential. The perfusion system was used to prepare the DUs, and the prepared DUs were systematically evaluated. The DU coating was prepared by enzymatic digestion for cell culture. The cASCs were seeded on the coverslips covered with DU coating and samples were collected on days 3, 7, and 10. Immunofluorescence staining and molecular biology testing were used to examine the differentiation of cASCs seeded on the DU coating. The cASCs were isolated and identified by flow cytometry. The prepared DUs removed the nuclear materials, and the 3-dimensional structure and biological compositions of the ureter were well preserved. Immunofluorescence staining showed the expression of anti-alpha smooth muscle actin (α-SMA). Western blot results suggested that the content of α-SMA in the experimental group was significantly higher than that in the control group at 3 different time points, and the mRNA expressions of α-SMA in the experimental group gradually increased with extended the culture time, whereas there was no significant change in the control group. The cASCs seeded on the coverslips of DU coating could differentiate into smooth muscle cells, and the number of differentiated cASCs increased significantly with extended incubation time.