147 DIFFERENTIATION OF HUMAN BONE MARROW STROMAL CELLS INTO AN UROEPITHELIAL LINEAGE

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2010147 DIFFERENTIATION OF HUMAN BONE MARROW STROMAL CELLS INTO AN UROEPITHELIAL LINEAGE Elena McNeill, Shantaram Bharadwaj, Anthony Atala, and Yuanyuan Zhang Elena McNeillElena McNeill More articles by this author , Shantaram BharadwajShantaram Bharadwaj More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and Yuanyuan ZhangYuanyuan Zhang More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.200AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The current strategy for engineering bladder tissues for reconstruction of the urinary tract involves obtaining a biopsy from patients for use in in vitro cell expansion. However, this approach cannot be used in bladder cancer patients requiring a complete bladder replacement. Bone marrow stem cells (BMSC) are capable of differentiating into multiple cell types. The aim of this study was to determine whether co-culture with human urothelium or culture in urothelium-conditioned medium (CM) could induce human BMSC to differentiate into urothelial like cells. These differentiation techniques could provide a novel source of urothelial cells for bladder tissue engineering in patients with bladder cancer. METHODS Human BMSC were purchased from a commercial source (Lonza). Flow cytometry was used to confirm that BMSC expressed the mesenchymal stem cell markers. Urothelium were obtained from bladder or ureter biopsy samples and cultured in keratinocyte serum-free medium. For co-culture experiments, BMSC were cultured with urothelium in a transwell plate. Urothelium were seeded on top of the permeable membrane within each well of this plate, and BMSC were cultured on a glass cover-slip beneath the membrane. For studies using the CM, media was collected every 12 hours from urothelium cultures maintained at 70% confluence. BMSC were seeded on glass cover-slips and placed on the bottom of 12-well plates. Several growth factors (PDGF-BB, VEGF, EGF, TGF-beta, bFGF and HGF) in urothelim CM were measured with ELISA. BMSC were cultured in the CM for urothelial differentiation. Cells were processed on days 3, 7, and 14 after seeding for RT-PCR and immunefluorescence analysis of uroplakin Ia, a specific urothelial cell markers, and expression of a panel of cytokeratins (CytoPan). RESULTS Prior to any treatment, BMSC did not express urothelial markers. After being co-cultured with UC or cultured using UC-derived CM, human BMSC expressed UC-specific genes and proteins: uroplakin-Ia, cytokeratin-7 and cytokeratin-13. PDGF-BB and VEGF were detected in the co-cultured or conditioned media. Urothelial differentiation of BMSC displayed time-dependent manner. Uroepithelial differentiation efficiency of BMSC was about 50% using co-culture and CM for up to 14 days. CONCLUSIONS This study demonstrated that human BMSC are able to give rise to bladder epithelial like cells when exposed to UC-conditioned media. BMSC could be an alternative cell source for urothelial differentiation in urinary tract reconstruction for cancer patients who need bladder replacement. Winston-Salem, NC© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e60 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Elena McNeill More articles by this author Shantaram Bharadwaj More articles by this author Anthony Atala More articles by this author Yuanyuan Zhang More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...