A model of superficial bladder cancer using fluorescent tumour cells in an organ-culture system.
OBJECTIVE
To develop a reproducible in vitro simulation of superficial bladder cancer for testing cytotoxic agents at clinically relevant concentrations.


METHODS
Square explants (5 mm) of rat bladder were cultured in Petri dishes in minimal volumes of Waymouth's MB 752/1 medium supplemented with 10% fetal calf serum, antibiotics and glutamine. Parental and resistant MGH-U1 urothelial cancer cells were transfected with a green fluorescent protein (GFP) vector. Transfectants were purified by flow cytometry. Cells were seeded onto the prepared organ cultures and images obtained using confocal microscopy. The tumour colonies were confirmed using scanning electron microscopy. Conventional intravesical cytotoxic agents including epirubicin, mitomycin-C and estramustine were tested in the system.


RESULTS
Colonies of GFP-MGH-U1 cells became established on the explants and were identified by confocal microscopy; the development of the colonies was then followed over several days. Staining the explant for viability allowed imaging of normal urothelium on the explant surface or surrounding skirt of urothelial cells. The conventional cytotoxic agents epirubicin, mitomycin C and estramustine showed the expected differential responses to parental and resistant cell types. The colonies were able to survive high concentrations of the drug, equivalent to those in clinical use. The colonies were imaged serially over a period of several days.


CONCLUSIONS
This system provides a more realistic model for testing cytotoxic agents for use in intravesical therapy, by allowing clinically relevant concentrations of drugs to be tested. The differential properties of the parental and resistant cells are maintained. The model also enables the same tumour colony to be imaged over several days in culture. The model may also be adapted for use in testing the effects of drugs on normal urothelium and the study of the effects of growth factors.
Crook
T J
TJ
MDR Research Group, Department of Urology, Southampton General Hospital, Southampton, UK. timjcrook@chandlers69.freeserve.co.uk
Hall
I S
IS
Solomon
L Z
LZ
Birch
B R
BR
Cooper
A J
AJ
D000818:Animals; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis; D023421:Models, Animal; D008954:Models, Biological; D009924:Organ Culture Techniques; D051381:Rats; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins