title: Investigations of urothelial cells seeded on commercially available small intestine submucosa.
abstract: OBJECTIVE
  To examine adherence and viability of human urothelial cells seeded on commercially available small intestine submucosa (SIS) specimens under serum-free conditions.
  
  
  METHODS
  Before seeding, SIS was either washed with incubation medium or coated with collagen A, fibronectin, or pronectin. A possible influence of SIS itself on the viability of urothelial cells was analysed with conditioned cell culture medium obtained by incubation of SIS for 24hours. In addition, untreated SIS and a setting without SIS were used as controls. Viability of urothelial cells was analysed with the WST-1 assay until day 9. Histology of seeded and unseeded SIS specimens was investigated after Papanicolaou staining. To demonstrate urothelial cell adherence on SIS, immunohistology was performed with a mixture of monoclonal AE1 and AE3 anticytokeratin antibodies.
  
  
  RESULTS
  Urothelial cells seeded on SIS revealed no measurable cell viability. SIS-conditioned cell culture medium was cytotoxic for urothelial cells after 24 hours. Histology only demonstrated cell nuclei and no cytoplasm both in seeded and unseeded SIS specimens, thus indicating porcine DNA. Expression of the cell type-specific marker proteins AE1/AE3 could not be demonstrated.
  
  
  CONCLUSIONS
  Since the commercially available SIS specimens used contained porcine DNA residues and demonstrated cytotoxic effects on urothelial cells, SIS is not suitable for in vitro construction of urothelial cell-matrix implants.
authors.affiliation: Department of Urology, University of Tübingen, Tübingen, Germany. gerhard.feil@med.uni-tuebingen.de <gerhard.feil@med.uni-tuebingen.de>
authors.forename: Gerhard
authors.initials: G
authors.lastname: Feil
authors.forename: Margot
authors.initials: M
authors.lastname: Christ-Adler
authors.forename: Sabine
authors.initials: S
authors.lastname: Maurer
authors.forename: Stefan
authors.initials: S
authors.lastname: Corvin
authors.forename: Hans-Oliver
authors.initials: HO
authors.lastname: Rennekampff
authors.forename: Jutta
authors.initials: J
authors.lastname: Krug
authors.forename: Jörg
authors.initials: J
authors.lastname: Hennenlotter
authors.forename: Ursula
authors.initials: U
authors.lastname: Kuehs
authors.forename: Arnulf
authors.initials: A
authors.lastname: Stenzl
authors.forename: Karl-Dietrich
authors.initials: KD
authors.lastname: Sievert
mesh_terms: D002448:Cell Adhesion / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000166:cytology*; D007421:Intestine, Small / Q000166:cytology*; D019459:Urothelium / Q000166:cytology*
chemical_list: D017077:Culture Media, Conditioned