title: Characterization of neuropathic bladder smooth muscle cells in culture.
abstract: OBJECTIVE
  Clinically bladder cells used in tissue engineering techniques will come from neuropathic bladders and not normal bladders. We determined if neuropathic bladder smooth muscle (SM) cells (SMCs) retain functional differences when cultured in vitro.
  
  
  METHODS
  Primary cultures of SMCs were established from patients with a neuropathic bladder (5) and a normal bladder (5). Expression of alpha-SM actin and SM myosin heavy chain was determined using immunocytochemical staining and Western blot analysis. Baseline cell proliferation and the mitogenic response to angiotensin II was assessed by cell counting and cell viability assays. Cell contractility was determined for normal and neuropathic SMCs using an in vitro collagen lattice assay. Cell adherence was measured assessed using partial and complete trypsinization assays.
  
  
  RESULTS
  Normal and neuropathic SMCs showed similar morphology in culture, and similar patterns of alpha-SM actin and SM myosin expression. Following 10 days of plating under optimal growth conditions the number of neuropathic SMCs was 170% more than normal SMCs. In response to angiotensin II neuropathic SMCs reached 54% of maximal growth capacity as opposed to 30% for normal SMCs (p <0.01). Neuropathic SMCs contracted significantly less in 10% serum and calcium ionophore (p <0.05), as determined by in vitro contractility assays. Neuropathic SMCs had 19% and 30% less adherent cells than normal SMCs (p <0.01) following isotonic solution washes and trypsinization, respectively.
  
  
  CONCLUSIONS
  These results demonstrate that cultured neuropathic bladder SMCs possess and maintain different characteristics than normal SMCs in vitro. The potential clinical implications of using these cells in conjunction with tissue engineering techniques for the promotion of bladder regeneration requires further investigation.
authors.affiliation: Department of Urology and Cell Biology, Children's Hospital of Oklahoma and University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA.
authors.forename: Hsueh-Kung
authors.initials: HK
authors.lastname: Lin
authors.forename: Rick
authors.initials: R
authors.lastname: Cowan
authors.forename: Pete
authors.initials: P
authors.lastname: Moore
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Qing
authors.initials: Q
authors.lastname: Yang
authors.forename: John A
authors.initials: JA
authors.lastname: Peterson
authors.forename: James J
authors.initials: JJ
authors.lastname: Tomasek
authors.forename: Bradley P
authors.initials: BP
authors.lastname: Kropp
authors.forename: Earl  Y
authors.initials: E
authors.lastname: Cheng
mesh_terms: D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D006801:Humans; D009130:Muscle, Smooth / Q000473:pathology*; D001750:Urinary Bladder, Neurogenic / Q000473:pathology*