[A New Ex-Vivo 3D Bladder Matrix Model for Standardised Evaluation of Urothelial Transfection Methods].
OBJECTIVE
Gene therapy strategies are a promising new alternative option in the treatment of cancer diseases and great effort is dedicated to the development of new gene transfer methods. At present, in vitro cell culture experiments or in vivo animal trials are the only available alternatives in the search for new gene transfer methods. We attempted to develop and evaluate a new 3D matrix model as a step between in vitro experiments and animal trials.


METHODS
We generated a convenient model with an agarose gel as a basis for small intestinal submucosa or a polyethylene membrane. The produced model consisting of human smooth muscle cells and human bladder carcinoma cells was transfected with a modified standard Lipofectamine trade mark 2000 transfection procedure and visualised by fluorescence microscopy after cryo-sectioning.


RESULTS
With the help of this new technique it is possible to generate three dimensional tissues consisting of different types of cells in which the cells are adherent on the polyethylene and the SIS-membrane during the entire treatment. The resulting model was successfully transfected with the pEGFP-N1 plasmid.


CONCLUSIONS
This new three dimensional model allows the standardised evaluation of new transfection methods on multilayered ex-vivo generated tissues consisting of different cells.
Schaaf
A
A
Urologische Klinik des Universitätsklinikums Mannheim, Mannheim, Deutschland.
McNealy
T
T
Steidler
A
A
Alken
P
P
Michel
M S
MS
D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003470:Culture Media; D015653:Cystectomy; D015316:Genetic Therapy; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D008055:Lipids / Q000494:pharmacology; D008081:Liposomes; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008961:Models, Structural; D009130:Muscle, Smooth / Q000166:cytology; D013997:Time Factors; D023822:Tissue Engineering; D014162:Transfection / Q000379:methods*; D001743:Urinary Bladder / Q000166:cytology*; D001749:Urinary Bladder Neoplasms; D019459:Urothelium / Q000166:cytology
D003470:Culture Media; D007202:Indicators and Reagents; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes