An Orthotopic Model of Murine Bladder Cancer

Summary

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

Abstract

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Protocol

1. Animals

1. Female C57BL/6J mice (Mus musculus) are verified free of contagious ectoparasites, helminth endoparasites, and antibodies to 17 murine viruses by the vendor prior to shipping. Mice are housed in the AAALAC-accredited animal facility of Tulane University in accordance with Guide for the Care and Use of Laboratory Animals.¹
2. Specifically, mice are group housed (4-5 mice/cage) in individually ventilated polycarbonate cageswith microisolation filter tops on hardwood bedding. They are provided ad libitum pelleted rodent food and tap water.
3. Mice are allowed 7 days to acclimate before the start of t....

Discussion

This minimally invasive method of establishing orthotopic bladder tumors in female mice can also be applied for cell types other than MB49, although differing results should be anticipated. When less-aggressive tumor cells are used, increasing the wattage slightly (using 0.5 W increments to establish the new optimum) will yield an area more amenable to tumor cell attachment, and longer bladder incubation times for the instilled cells will yield a greater amount of cell attachment at the burn sites (unpublished data). A.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
C57Bl/6J mice	Jackson Laboratory	000664	3-5-wk-old
Polycarbonate cages and ventilated rack	Lab Products	Super Mouse 750	327mm x 190mm x 143mm
Hardwood Bedding	Harlan Laboratories	Laboratory –grade SaniChips
Rodent diet	Harlan Laboratories	Rodent Diet 2019
IV Catheter	MWI Veterinary Supply	009231	24 G x ¾ in.
Electrocautery unit	Bovie Medical Corporation	Aaron 900
Ultrasound unit	Sonosite	180 Plus (Vet)
MB49 cell line	Wake Forest University Baptist Medical Center	N.A.
DMEM	Invitrogen	21063-029
Fetal Bovine Serum	Gemini Bio Products	100106 500A57B
Penicillin-Streptomycin	Invitrogen	15140-163