Investigations of urothelial cells seeded on commercially available small intestine submucosa.
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.
Feil
Gerhard
G
Department of Urology, University of Tübingen, Tübingen, Germany. gerhard.feil@med.uni-tuebingen.de <gerhard.feil@med.uni-tuebingen.de>
Christ-Adler
Margot
M
Maurer
Sabine
S
Corvin
Stefan
S
Rennekampff
Hans-Oliver
HO
Krug
Jutta
J
Hennenlotter
Jörg
J
Kuehs
Ursula
U
Stenzl
Arnulf
A
Sievert
Karl-Dietrich
KD
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*
D017077:Culture Media, Conditioned