title: A modified method by differential adhesion for enrichment of bladder cancer stem cells.
abstract: OBJECTIVE
  In a previous study the vaccine was effective against bladder cancer in a mouse model. However, a small portion of tumors regrew because the vaccine could not eliminate bladder cancer stem cells (CSCs). In this study, we showed a modified method for the isolation of bladder CSCs using a combination of differential adhesion method and serum-free culture medium (SFM) method.
  
  
  METHODS
  Trypsin-resistant cells and trypsin-sensitive cells were isolated from MB49, EJ and 5637 cells by a combination of differential adhesion method and SFM method. The CSCs characterizations of trypsin-resistant cells were verified by the flow cytometry, the western blotting, the quantitative polymerase chain reaction, the resistance to chemotherapy assay, the transwell assay, and the tumor xenograft formation assay.
  
  
  RESULTS
  Trypsin-resistant cells were isolated and identified in CSCs characters, with high expression of CSCs markers, higher resistance to chemotherapy, greater migration in vitro, and stronger tumorigenicity in vivo.
  
  
  CONCLUSIONS
  Trypsin-resistant cells displayed specific CSCs properties. Our study showed trypsin-resistant cells were isolated successfully with a modified method using a combination of differential adhesion method and SFM method.
authors: Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China.|Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors: Yong-Tong
authors: YT
authors: Zhu
authors: Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors: Shi-Yu
authors: SY
authors: Pang
authors: Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors: Yang
authors: Y
authors: Luo
authors: Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors: Wei
authors: W
authors: Chen
authors: Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors: Ji-Ming
authors: JM
authors: Bao
authors: Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
authors: Wan-Long
authors: WL
authors: Tan
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor; D019496:Cancer Vaccines / Q000276:immunology; D002448:Cell Adhesion / Q000187:drug effects*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D016895:Culture Media, Serum-Free; D005434:Flow Cytometry; D051379:Mice; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000166:cytology*; D060888:Real-Time Polymerase Chain Reaction; D014357:Trypsin / Q000494:pharmacology*; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
chemical_list: D014408:Biomarkers, Tumor; D019496:Cancer Vaccines; D016895:Culture Media, Serum-Free; D014357:Trypsin
keywords: Neoplastic Stem Cells; Trypsin; Urinary Bladder Neoplasms