Self-Renewal and Differentiation Capacity of Urine-Derived Stem Cells after Urine Preservation for 24 Hours
Figure 5
Urothelial differentiation of USCs.
(A) Preserved USCs and fresh USCs 14 days after urothelial differentiation expressed urothelial cell markers (uroplakin Ia/III, CK7, CK13 and CK20) that were similar to the urothelial cells, while few uninduced USCs expressed these markers assessed by immunofluorescent staining. Scale bar = 50 uM. (B) Preserved USCs and fresh USCs expressed almost same amount of urothelial-specific proteins (uroplakin Ia/III, CK7, E-cadherin and cingulin) assessed with Western blotting. (C) Preserved USCs and fresh USCs displayed tight junction markers (cingulin and E-cadherin) on cell membrane boundaries (arrow) between cells detected by immunofluorescent staining. Scale bar = 20 uM. (D) Preserved USCs and fresh USCs exhibited tight junction-desmosomes (arrows) between two adjacent cells, whereas non-induced fresh USCs and preserved USCs showed desmosomes when examined by transmission electron microscopy (TEM). (E) A barrier function assay performed by using a fluorescent tracer on confluent cells cultured on inserts. Both preserved USCs and fresh USCs showed similar leakage protection that was significantly better than uninduced USCs (p<0.05).