172 IN VIVO SKELETAL MYOGENIC DIFFERENTIATION OF AUTOLOGOUS URINE-DERIVED STEM CELLS FOR POTENTIAL APPLICATION IN TREATMENT OF URINARY INCONTINENCE

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2011172 IN VIVO SKELETAL MYOGENIC DIFFERENTIATION OF AUTOLOGOUS URINE-DERIVED STEM CELLS FOR POTENTIAL APPLICATION IN TREATMENT OF URINARY INCONTINENCE Shantaram Bharadwaj, GuiHua Liu, Shaofeng Wu, Anthony Atala, and Yuanyuan Zhang Shantaram BharadwajShantaram Bharadwaj 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.2011.02.241AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Autologous adult stem cell injection therapy has recently provided a promising alternative for sphincter tissue regeneration and treatment of stress urinary incontinence (SUI). Even though stem cells obtained from skeletal muscle, bone marrow, and adipose tissues are regarded as possible candidates for use in this therapy, their harvest involves an invasive procedure and may cause complications. Obtaining an autologous stem cell source non-invasively would be highly desirable. Recently, we have established a primary culture system to isolate and repopulate stem cells from voided human urine. These cells, called urine-derived stem cells (USC), possess extensive expansion capability and multipotent differentiation potential. USC can also differentiate into mesodermal cell lineages, including skeletal myocytes, in vitro. The goal of this study was to determine if USC could differentiate into skeletal muscle cells in vivo for potential use in cell therapies for SUI. METHODS Urine derived cells were harvested from healthy individuals (n=9), characterized with perictye/mesenchymal stem cell surface markers and then induced to differentiate into skeletal muscle cells with myogenic differentiation media. The cells were cultured for up to 2 weeks and were compared to cells cultured in medium conditioned with skeletal muscle cells. Induced cells were assessed for alterations in morphology and expression of skeletal muscle cell markers (MyoD, Myogenin, Myosin, Desmin, M-Cadherin and Myf5) via RT-PCR, Western blot and immunefluorescence. Differentiated (n=4) and non-differentiated USC (n=4, control) (1 million in 100 μl) were injected into the leg muscle of athymic mice. These muscle tissues were harvested 4 weeks after cell injection and assessed using histological and immunohistochemical methods (triple staining for DAPI, human nuclei and skeletal muscle markers). RESULTS Injected cells could be identified in the host muscle tissue. Both skeletal muscle-differentiated USC and non-differentiated USC expressed human nuclear markers and the skeletal muscle proteins (myoD, myogenin, myosin, desmin, M-Cadherin, and Myf5). However, more cells expressed both types of protein markers in the differentiated USC group compared to those in the non-differentiated USC group. CONCLUSIONS USC can differentiate into skeletal muscle cells in vivo after pre-conditioning with specific differentiation media. USC may be a potential source of cells for injection therapy in the treatment of urethral sphincter dysfunction. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e72 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shantaram Bharadwaj 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 ...