1609 POTENTIAL USE FOR UROTHELIAL MUCOSA ENGINEERED WITH AUTOLOGOUS URINE DERIVED STEM CELLS SEEDED ON A POROUS COLLAGEN MATRIX IN HYPOSPADIAS REPAIR

Abstract

You have accessJournal of UrologyPediatrics: Congenital Anomalies - Lower Urinary Tract & Genitalia II1 Apr 20121609 POTENTIAL USE FOR UROTHELIAL MUCOSA ENGINEERED WITH AUTOLOGOUS URINE DERIVED STEM CELLS SEEDED ON A POROUS COLLAGEN MATRIX IN HYPOSPADIAS REPAIR Rongpei Wu, Guihua Liu, Shaofeng Wu, Anthony Atala, and Yuanyuan Zhang Rongpei WuRongpei Wu Winston Salem, NC More articles by this author , Guihua LiuGuihua Liu Winston Salem, NC More articles by this author , Shaofeng WuShaofeng Wu Winston Salem, NC More articles by this author , Anthony AtalaAnthony Atala Winston Salem, NC More articles by this author , and Yuanyuan ZhangYuanyuan Zhang Winston Salem, NC More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1404AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Surgical repair of hypospadias often requires autogenous tissue, such as foreskin, bladder or oral mucosa to extend urethral segments. However, the procedure required to obtain these tissues is an invasive approach and it may lead to donor site morbidity. Additionally, adequate tissue for this substitution is not always available, and if the urethral defect is too long, this type of repair is not possible. Tissue-engineered tubular urethral tissues are an alternative option and allow for replacement of deficient urethral tissues with functionally equivalent ones. The goal of this study was to generate a 3-dimensional porous small intestinal submucosa (SIS) scaffold and recellularize this scaffold with urothelial cells (UC) and smooth muscle cells (SMC) derived from the differentiation of urine-derived stem cells (USC) in order to form engineered urethral tissue that could be used in hypospadias repair. METHODS Fresh SIS derived from pigs was decellularized with 5% peracetic acid (PAA). Cells were obtained from voided urine samples from healthy individuals. USC were isolated, characterized and induced to differentiate into UC and SMC. Differentiated UC and SMC derived from USC were seeded onto the SIS scaffolds in a layered co-culture fashion and cultured under dynamic conditions for one week. The seeded cells formed multiple uniform layers on the SIS and penetrated deeper into the porous matrix during dynamic culture. Urothelial and muscle cells derived from USC expressed UC markers (Uroplakin-III, AE1/AE3) or SMC markers (α-SM actin, desmin, and myosin), respectively, after implantation into athymic mice. RESULTS DNA content analysis showed that most nuclear material was removed from the SIS scaffold. Treatment with 5% PAA led to development of a highly porous surface with retention of less cellular material, and the decellularized SIS maintained the majority of the tensile strength of normal tissue. In 3-D dynamic culture, the seeded cells (UC and SMC) formed multiple, even layers on the surface of matrix. Cells also penetrated deeper into the lamina propria of the matrix compared to untreated matrix. Immunocytochemical staining indicated that the grafted USC expressed urothelial- and smooth muscle-specific markers both, in vitro and in vivo. CONCLUSIONS This study demonstrates that the decellularized SIS possesses appropriate 3-D porosity to prompt the induced USC to form multilayer of urothelium and SMC-matrix infiltration, which provides potential applications in urethral reconstruction in hypospdias repair. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e651 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rongpei Wu Winston Salem, NC More articles by this author Guihua Liu Winston Salem, NC More articles by this author Shaofeng Wu Winston Salem, NC More articles by this author Anthony Atala Winston Salem, NC More articles by this author Yuanyuan Zhang Winston Salem, NC More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...