Laparoscopy in Ureteral Engineering: A Feasibility Study Hervé Baumert, Mehrak Hekmati, Irene Dunia, Dhouha Mansouri, Walid Massoud, Vincent Molinié, E. Lucio Benedetti, Bernard Malavaud  European Urology  Volume 54, Issue 5, Pages 1154-1163 (November 2008) DOI: 10.1016/j.eururo.2008.01.012 Copyright © 2008 Terms and Conditions

Fig. 1 (A) Port positioning during the bladder harvesting; (B–D) laparoscopic bladder harvest using a 45-mm endo GIA stapler; (E) bladder specimen after resection of the stapling line. The urothelial cells were scraped away from the bladder wall with a knife blade. European Urology 2008 54, 1154-1163DOI: (10.1016/j.eururo.2008.01.012) Copyright © 2008 Terms and Conditions

Fig. 2 (A) Port positioning during the laparoscopic transfer of the construct in the omentum. (B) The seeded scaffold is shaped around a 20-Fr silicone drain, then introduce into the abdominal cavity through a 10-mm port. The construct is wrapped by the omentum (C) and then fixed to it using 3/0 interrupted prolene sutures (D, E). European Urology 2008 54, 1154-1163DOI: (10.1016/j.eururo.2008.01.012) Copyright © 2008 Terms and Conditions

Fig. 3 (A) The bladder 2 wk after harvest. No adhesion was observed, even in front of the stapling line. (B, C) Laparoscopic evaluation after the omental maturation of the construct. Again, no adhesion or inflammatory response was observed, and the construct remained freely mobile. (D) Open harvest of the construct. Note the dense vascular supply from the omentum. (E) Aspect after formalin fixation of the engineered ureter and its omental pedicle. European Urology 2008 54, 1154-1163DOI: (10.1016/j.eururo.2008.01.012) Copyright © 2008 Terms and Conditions

Fig. 4 (A) The engineered ureter and its omental pedicle after H&E staining. (B–E) High-power views (250×) of the construct after omental maturation: (B) H&E staining reveals a multilayered structure with no evidence of inflammatory reaction; (C) anti-cytokeratin 7 immunostaining confirms the presence of a differentiated multilayer urothelium; (D) the urothelium is formed over connective tissue composed of smooth muscle cells with α-actin smooth muscle immune reactivity. (E) Anti–Von Willebrand factor immunostaining shows large vessels and a dense capillary network. European Urology 2008 54, 1154-1163DOI: (10.1016/j.eururo.2008.01.012) Copyright © 2008 Terms and Conditions

Fig. 5 (A) Superficial layer, close to the silicone drain of the neoureter, composed of highly polarized elongated cells with the apical surface characterized by raised ridges and flat areas. The cells are associated by tight junctions (arrows), and the cell layer is supported by collagen fibres and a basal membrane (6000×). (B) Intermediate zone of the neoureter comprising smooth fibre cells characterized by intermingled filamentous bundles and cytoplasmic condensations, which are particularly evident at the inner aspect of the plasmalemma (arrows). In the smooth fibre cell cytoplasm, large areas are occupied by the endoplasmic reticulum (ER) and polyribosomes. Collagen fibres (CF) appear in the intercellular space (12,000×). (C) High magnification (25,000×) of a smooth muscle cell showing a great number of caveloae in close association with the plasma membrane. Arrows point to cytoplasmic condensations crossing the filamentous bundles. (D) The inner neoureter zone, close to the omentum (Om). Fibroblast and CFs are present. The fibroblast cytoplasm contains mitochondria and ER profiles. The omentum cells are characterized by large fat deposits outlined by a thin cytoplasmic layer (8000×). European Urology 2008 54, 1154-1163DOI: (10.1016/j.eururo.2008.01.012) Copyright © 2008 Terms and Conditions

Fig. 6 (A) Urothelial cell characterized by the presence of a discoid fusiform vesicle. The arrow points to microtubules, and arrowheads point to basal membrane (70,000×). (B) High magnification (450,000×) of a membrane profile outlined by a rectangle in (A). The membrane displays an unusual subunit organization suggestive of a planar uroplakin(s) crystal to be inserted into the asymmetric unit membrane. (C) High magnification (180,000×) of the urothelium surface, where the plasma membrane is characterized by an asymmetric unit membrane with a thick and electron-dense outer layer (arrow). The cytoplasmic vesicles, close to the inner aspect of the asymmetric unit membrane, display the same asymmetric membrane bilayer (arrows). (D) High magnification (180,000×) of a region of close contact between two cell surfaces. Note the presence of a tight junction (arrowhead) and the asymmetric features of the plasma membrane bilayers (arrow). European Urology 2008 54, 1154-1163DOI: (10.1016/j.eururo.2008.01.012) Copyright © 2008 Terms and Conditions