BACKGROUND: The purpose of the present study was to assess the differentiation potential of adipose-derived stem cells (ASCs) into smooth muscle cells (SMCs) and their potential for promoting regeneration of smooth muscle for ureteral tissue engineering. METHODS: ASCs were isolated, proliferated, and identified in vitro. SMC differentiation was induced using SMC induction medium. Gene expression was evaluated by quantitative polymerase chain reaction, immunofluorescence, and Western blotting. Vessel extracellular matrix was obtained by a decellularization process. The induced cells were seeded onto vessel extracellular matrix for ureter reconstitution. Grafts were obtained for evolutionary histologic studies. Renal function and ureteral patency was evaluated by intravenous urography at 16 wk. RESULTS: Flow cytometry demonstrated that the ASCs expressed CD90, but did not express CD45 or CD34. After 6 wk of induction, upregulation of α-smooth muscle actin expression was determined by quantitative polymerase chain reaction, and smooth muscle myosin heavy chain expression was confirmed by immunofluorescence and Western blotting in the induced cells. Vessel extracellular matrix exhibited a nontoxic and bioactive effect on the induced cells. Histologically, stratified urothelium and organized muscle bundles were observed in the grafts at 16 wk. Intravenous urography demonstrated no ureteral stricture or hydroureteronephrosis. CONCLUSIONS: These results have demonstrated that ASCs can be differentiated into SMCs and this potential promoted smooth muscle regeneration for ureteral tissue engineering.
BACKGROUND: The purpose of the present study was to assess the differentiation potential of adipose-derived stem cells (ASCs) into smooth muscle cells (SMCs) and their potential for promoting regeneration of smooth muscle for ureteral tissue engineering. METHODS: ASCs were isolated, proliferated, and identified in vitro. SMC differentiation was induced using SMC induction medium. Gene expression was evaluated by quantitative polymerase chain reaction, immunofluorescence, and Western blotting. Vessel extracellular matrix was obtained by a decellularization process. The induced cells were seeded onto vessel extracellular matrix for ureter reconstitution. Grafts were obtained for evolutionary histologic studies. Renal function and ureteral patency was evaluated by intravenous urography at 16 wk. RESULTS: Flow cytometry demonstrated that the ASCs expressed CD90, but did not express CD45 or CD34. After 6 wk of induction, upregulation of α-smooth muscle actin expression was determined by quantitative polymerase chain reaction, and smooth muscle myosin heavy chain expression was confirmed by immunofluorescence and Western blotting in the induced cells. Vessel extracellular matrix exhibited a nontoxic and bioactive effect on the induced cells. Histologically, stratified urothelium and organized muscle bundles were observed in the grafts at 16 wk. Intravenous urography demonstrated no ureteral stricture or hydroureteronephrosis. CONCLUSIONS: These results have demonstrated that ASCs can be differentiated into SMCs and this potential promoted smooth muscle regeneration for ureteral tissue engineering.