title: Isolation and characterization of smooth muscle cells from rat corpus cavernosum tissue for the study of erectile dysfunction.
abstract: OBJECTIVE
  Primary culture of the cavernous smooth muscle cells from corpus cavernous tissues is known to be difficult, mainly because of contamination with fibroblasts. We applied a new method for better isolation of rat penile smooth muscle cells (RPSMCs) from rat corpus cavernosum tissue for reliable ex vivo research on erectile dysfunction.
  
  
  METHODS
  With the use of 8-week-old adult male Sprague-Dawley rats, ex vivo migrations of rat cavernous tissue were measured by penis and aortic ring assay by use of a Matrigel-based D-valine-modified culture method. The expression of α-smooth muscle actin (α-SMA) and platelet/endothelial cell adhesion molecule (PECAM)-1 in the RPSMCs was determined by standard immunofluorescent staining and immunoblotting. The expression patterns of phosphodiesterase (PDE) family mRNA in RPSMCs were compared with patterns in rat aortic smooth muscle cells (RASMCs) by use of quantitative real-time reverse transcription polymerase chain reaction.
  
  
  RESULTS
  Immunocytochemical staining showed greater α-SMA-positive and PCAM-1-negative fluorescence. Moreover, whereas the expression of α-SMA was detected in the RPSMCs, that of PECAM-1 was not. The levels of PDE1A, PDE1B, PDE1C, PDE2A, PDE3A, PDE4A, PDE4B, PDE4C, PDE4D, and PDE5A mRNA in the RPSMCs were about 3.2-, 4.4-, 3.4-, 29.0-, 3.5-, 2.8-, 2.9-, 6.1-, 45.0-, and 6.0-fold the corresponding expression in RASMCs.
  
  
  CONCLUSIONS
  We developed a two-stage tissue culture method utilizing a Matrigel-based sprouting culture system to facilitate stromal cell sprouting and an adherent culture system using D-valine to eliminate the contamination of fibroblasts into the smooth muscle cells.
authors.affiliation: Department of Urology, Konkuk University School of Medicine, Chungju, Korea.
authors.forename: Hong
authors.initials: H
authors.lastname: Chung
authors.forename: Seung-Hyo
authors.initials: SH
authors.lastname: Jung
authors.forename: Ji-Kan
authors.initials: JK
authors.lastname: Ryu
authors.forename: Bokyung
authors.initials: B
authors.lastname: Kim
authors.forename: Hong Sup
authors.initials: HS
authors.lastname: Kim
authors.forename: Sang-Kuk
authors.initials: SK
authors.lastname: Yang