Volume 45, Issue 6, Pages 799-805 (June 2004) Growth, Differentiation and Senescence of Normal Human Urothelium in an Organ-Like Culture  Ahmad Daher, Willem I de Boer, Marie-Aude Le Frère-Belda, Laurence Kheuang, Claude C Abbou, François Radvanyi, Marie-Claude Jaurand, Jean Paul Thiery, Sixtina Gil Diez de Medina, Dominique K Chopin  European Urology  Volume 45, Issue 6, Pages 799-805 (June 2004) DOI: 10.1016/j.eururo.2004.01.002

Fig. 1 Comparison of cell spreading (a) of normal urothelium from ureter explant on membranes previously treated separately with matrix component: type IV collagen (•), fibronectin (○) laminin (▴) or BSA (□). (b) Comparison of cell spreading of normal urothelium from bladder (■) and ureter (▴) explants. European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)

Fig. 2 Percentage of cells incorporating BrdU (•) (left Y-axis) and number of cells per mm2 (bars; right Y-axis) between 0 and 30 days culture. European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)

Fig. 3 Ultrastructural features of differentiated normal urothelial cells on Cyclopore membranes after 20 days of culture: microvilli (open arrow) and glycocalyx (closed arrow) (a), desmosomes (asterisk) (a and b), asymmetric unit membrane (arrow) (b), rough endoplasmic reticulum (RER) (c), mitochondria (M) and Golgi apparatus (G) (d), intermediate filament (arrow) and vesicles (V) (e). European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)

Fig. 4 Immunohistochemistry showing cytokeratins CK17 (a–c) and CK18 (d–f) cytoplasm labeling on ureter sections (a,d) and in urothelium culture model after 10 (b,e) and 30 (c,f) days of culture, and BrdU nuclear staining (b,c,e,f) (bar=50μm). European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)

Fig. 5 Expression of mRNA transcripts of different markers of urothelial cells in time of culture. The expression is given as the ratio of CK7, CK18, or E-cadherin mRNA over TBP mRNA (a) and as the ratio of uroplakins UPIb and UPII mRNA over GAPDH mRNA (b) (significance level: p=0.001). European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)

Fig. 6 The expression of the mRNA transcripts of TGFα, HB-EGF and AR measured by semi-quantitative RT-PCR are represented as histograms. The expression is given as the ratio of factors mRNA over TBP mRNA. European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)

Fig. 7 (A) Level of p16/CDKN2A mRNA relative to the corresponding TBP mRNA level in the urothelium model at different times intervals as assessed by semi-quantitative RT-PCR. (B) Endogenous β-galactosidase detected after 10 (a), 15 (b), 20 (c), 25 (d), 30 (e) and 40 (f) days of culture. β-Galactosidase activity provided a blue staining of the cells that was not detected at the beginning (bar=50μm). European Urology 2004 45, 799-805DOI: (10.1016/j.eururo.2004.01.002)