Fig. 3

Characteristics and multipotent differentiation of USCs. a Single USCs can be expanded to a large population. Single cells appeared on day 4 (a a), clone formation was observed on day 6 and 7 (a bc), and cell confluency was achieved on day 11 (a d). Scale bar = 100 μm. b USCs (passage 2) were induced to differentiate into two distinct lineages. The morphology of USCs (b a) is shown after induction with SMC induction media (b c) and UC induction media (b b) for 14 days. Scale bar = 100 μm. c IF staining of myogenic-induced (c a–c c) and urothelial-induced (c d–c f) USCs. Specific marker staining (red) and nuclear staining by DAPI (blue) are shown. Scale bar = 50 μm. d Detection of the SMC-specific marker (myosin; d a) and UC-specific markers (AE1/AE3; d b) by Western blotting. Specific bands are seen in lanes with proteins from induced USCs and the positive control. DAPI 4′,6-diamidino-2-phenylindole, SMC smooth muscle cell, UC urothelial cell, USC urine-derived stem cell