179 THE DYNAMIC BLADDER MICROENVIRONMENT GOVERNS SMOOTH MUSCLE DIFFERENTIATION OF SKIN DERIVED PRECURSOR CELLS: IMPLICATIONS FOR TISSUE REGENERATION

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2011179 THE DYNAMIC BLADDER MICROENVIRONMENT GOVERNS SMOOTH MUSCLE DIFFERENTIATION OF SKIN DERIVED PRECURSOR CELLS: IMPLICATIONS FOR TISSUE REGENERATION Cornelia Tolg, Shaalee Dworski, Jeff Biernaskie, Karen Aitken, Freda Miller, and Darius Bagli Cornelia TolgCornelia Tolg Toronto, Canada More articles by this author , Shaalee DworskiShaalee Dworski Toronto, Canada More articles by this author , Jeff BiernaskieJeff Biernaskie Toronto, Canada More articles by this author , Karen AitkenKaren Aitken Toronto, Canada More articles by this author , Freda MillerFreda Miller Toronto, Canada More articles by this author , and Darius BagliDarius Bagli Toronto, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.248AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Smooth muscle cell containing organs such as bladder are targeted by a variety of pathological conditions necessitating surgery or organ replacement. Currently, augmentation surgery or tissue engineering approaches are hampered by lack of functional smooth muscle cells. Multipotent skin derived progenitor cells (SKPs) can easily be isolated from adult skin and can be differentiated into a variety of cell lineages. Here we describe FBS induced smooth muscle cell differentiation of SKPs and effects of the dynamic bladder microenvironment on this differentiation. METHODS SKPs were isolated from adult rat skin and cultured as non-adherent spheres in EGF and FGF containing medium. Differentiation was induced with 15% FBS. Expression of smooth muscle cell specific genes was analyzed by IF and q-PCR. Bladders of adult rats were stretched ex vivo and organoids were isolated by collagen digestion. To investigate the effect of bladder microenvironment, SKPs were co-cultured with organoids or exposed to medium conditioned by ex vivo bladder organ cultures. SKPs differentiation was analyzed as above. RESULTS Skps cultured in presence of FBS strongly upregulate expression of smooth muscle marker such as smooth muscle actin, calponin, myocardin and myosin heavy chain compared to SKPs cultured in EGF/FGF containing medium as demonstrated by real time PCR and IF. FBS treated SKPs organize smooth muscle actin into cytoskeleton actin cables and contract collagen I gels, two characteristics of functional smooth muscle cells. To investigate how a dynamic microenvironment characterized by strain/relaxation cycles modifies SMC differentiation of SKPs, we differentiated SKPs in presence of bladder organoids or conditioned medium derived from stretched or relaxed bladders. Diffusible factors released from stretched bladders decreased SMC differentiation of SKPs. Inhibition of mTOR signaling by Rapamycin treatment revealed that FBS induced SMC differentiation of SKPs requires mTOR signaling. CONCLUSIONS These results suggest that SKPs could be used as SMC source for augmentation surgery and tissue engineering of hollow organs such as bladder. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e74 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Cornelia Tolg Toronto, Canada More articles by this author Shaalee Dworski Toronto, Canada More articles by this author Jeff Biernaskie Toronto, Canada More articles by this author Karen Aitken Toronto, Canada More articles by this author Freda Miller Toronto, Canada More articles by this author Darius Bagli Toronto, Canada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...