257 FATE OF STEM CELLS IN INJURY: IN-VIVO REAL TIME TRACKING OF MESENCHYMAL STEM CELLS (MSCS) IN A RAT MODEL OF STRESS URINARY INCONTINENCE (SUI)

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2013257 FATE OF STEM CELLS IN INJURY: IN-VIVO REAL TIME TRACKING OF MESENCHYMAL STEM CELLS (MSCS) IN A RAT MODEL OF STRESS URINARY INCONTINENCE (SUI) Zhina Sadeghi, Joseph Molter, Michael Kavran, Donald Lennon, Kerry O. Grimberg, Firouz Daneshgari, Arnold Caplan, Zhenghong Lee, Chris A. Flask, and Adonis Hijaz Zhina SadeghiZhina Sadeghi Cleveland, OH More articles by this author , Joseph MolterJoseph Molter Cleveland, OH More articles by this author , Michael KavranMichael Kavran Cleveland, OH More articles by this author , Donald LennonDonald Lennon Cleveland, OH More articles by this author , Kerry O. GrimbergKerry O. Grimberg Cleveland, OH More articles by this author , Firouz DaneshgariFirouz Daneshgari Cleveland, OH More articles by this author , Arnold CaplanArnold Caplan Cleveland, OH More articles by this author , Zhenghong LeeZhenghong Lee Cleveland, OH More articles by this author , Chris A. FlaskChris A. Flask Cleveland, OH More articles by this author , and Adonis HijazAdonis Hijaz Cleveland, OH More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1639AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Previous studies have shown that human MSCs (hMSCs) improved incontinence in a rat model of vaginal distension (VD). This study sought to track periurethrally injected hMSCs using in vivo bioluminescence imaging (BLI). METHODS Bone marrow-derived hMSCs were transfected with a triple-fusion imaging reporter gene fluc-mrfp-ttk, encoding firefly luciferase, monomeric red fluorescent protein, and truncated herpes simplex virus type 1 sr39 thymidine kinase. The cells were then double labeled with naturally occurring flourophore compound from the fungus Epicoccum nigrum (LavaCell®). One Sprague-Dawley rat underwent 4hr of VD followed by injection of double labeled hMSCs (1.5 million cells). Another age-matched control rat received double labeled hMSCs without VD. Animals were imaged immediately and daily after injection with BLI until the signal disappeared. Animals were then sacrificed and sections of the midurethral and anterior vaginal wall at the site of injection were processed and stained. Another group of rats underwent VD followed by local injection of PKH-labeled hMSC and were sacrificed 2 hours and 24 hours after hMSC injection (n=4 per group). Sections of the midurethra were prepared and imaged under immunofluorescent microscope. RESULTS BLI signal increased within 3d in the pelvis of VD rats injected with hMSC. Interestingly, rats that received hMSC in the absence of VD demonstrated immediate localization to the lungs (Figure). The signal disappeared 7 days post-injection in both animals. Staining for Human-specific Alu stain at 7 days did not demonstrate positive cells in the urethra or connective tissue around urethra. PKH-labeled cells 2 hours and 24 hours after local injection demonstrated positive preurethral signal that coalesced in the VD group at 24 hours. CONCLUSIONS Transplanted hMSCs home to injured tissues where they exert their effect and proliferate. Cells were not found at the injection site 1 week later. . Follow up studies are needed to determine the sequence of events contributing to distribution of MSCs at 1 week. LavaCell® fluorophore dye keeps the proliferation characteristics of hMSCs almost intact. Combination of techniques is needed to effectively track cells. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e105-e106 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Zhina Sadeghi Cleveland, OH More articles by this author Joseph Molter Cleveland, OH More articles by this author Michael Kavran Cleveland, OH More articles by this author Donald Lennon Cleveland, OH More articles by this author Kerry O. Grimberg Cleveland, OH More articles by this author Firouz Daneshgari Cleveland, OH More articles by this author Arnold Caplan Cleveland, OH More articles by this author Zhenghong Lee Cleveland, OH More articles by this author Chris A. Flask Cleveland, OH More articles by this author Adonis Hijaz Cleveland, OH More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...