Epithelial-Differentiated Adipose-Derived Stem Cells Seeded Bladder Acellular Matrix Grafts for Urethral Reconstruction: An Animal Model

• Published in: Tissue engineering. Part A 2014-02, Vol.20 (3-4), p.774-784
• Main Authors: Li, Hongbin, Xu, Yuemin, Xie, Hong, Li, Chao, Song, Lujie, Feng, Chao, Zhang, Qin, Xie, Minkai, Wang, Ying, Lv, Xiangguo
• Format: Article
• Language: English
• Subjects: Adipose Tissue - cytology; Animals; Biomarkers - metabolism; Bromodeoxyuridine - metabolism; Cell Differentiation; Cell Survival; Epithelial Cells - cytology; Extracellular Matrix - transplantation; Fluorescent Antibody Technique; Keratin-13 - metabolism; Male; Matrix; Models, Animal; Myosins - metabolism; Original; Original Articles; Rabbits; Reconstructive Surgical Procedures; Skin & tissue grafts; Stem Cell Transplantation; Stem cells; Stem Cells - cytology; Stem Cells - ultrastructure; Urethra - surgery; Urinary Bladder - transplantation
• ISSN: 1937-3341; 1937-335X
• DOI: 10.1089/ten.tea.2013.0122

The limited amount of available epithelial tissue is considered a main cause of the high rate of urethral reconstruction failures. The aim of this study was to investigate whether epithelial-differentiated rabbit adipose-derived stem cells (Epith-rASCs) could play a role of epithelium in vivo functionally and be a potential substitute of urothelium. Substitution urethroplasty was performed to repair an anterior urethral defect in male New Zealand rabbits using Epith-rASCs seeded bladder acellular matrix grafts (BAMGs) after 5-bromo-2′-deoxyuridine (BrdU) labeling, based on the in vitro epithelial induction system we previously described. Urethroplasty with cell-free BAMGs and with undifferentiated rASCs (Und-rASCs) seeded BAMGs were performed as controls. After surgery, a notable amelioration of graft contracture and recovery of urethral continuity were observed in the Epith-rASCs/BAMG group by retrograde urethrograms and macroscopic inspection. Immunofluorescence revealed that the BrdU-labeled Epith-rASCs/Und-rASCs colocalized with cytokeratin 13 or myosin . Consistent with the results of western blotting, at early postimplantation stage, the continuous epithelial layer with local multilayered structure was observed in the Epith-rASCs/BAMG group, whereas no significant growth and local monolayer growth profile of epithelial cells were observed in the BAMG and Und-rASCs/BAMG group, respectively. The results showed that Epith-rASCs could serve as a potential substitute of urothelium for urethral tissue engineering and be available to prevent lumen contracture and subsequent complications including recurrent stricture.