Modeling Orthotopic Bladder Tumor in Mouse Model: A Procedure for Intravesical Administration of Cancer Cells into Murine Bladder

Abstract

Source: Tham, S. M. et al. A Murine Orthotopic Bladder Tumor Model and Tumor Detection System. J. Vis. Exp. (2017)

In this video, we describe the intravesical procedure of delivering cancer cells into the bladder of a mouse model. This procedure helps generate an orthotopic bladder tumor model that can be used to develop therapeutic strategies.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Implanting the Tumor

NOTE: Each mouse is implanted with 1 x 10⁵ MB49-PSA cells in 50 µL of DMEM blank media in the bladder. Due to the dead space in the catheter, always prepare extra volume (at least 100 µL extra per mouse). An alternative approach would be to use an air-filled syringe.

1. Cells
   1. One day before implantation, when the cells are approximately 80% confluent, passage the MB49....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
MB49-PSA cells			Ref (Wu QH, 2004)
Dulbecco's Modified Eagle's Medium (DMEM) media	Biowest	L0102
Fetal Bovine Serum (FBS) South American	Biowest	S1810
Fetal Bovine Serum (FBS) South American, Premium	Biowest	S181B
Fetal Bovine Serum (FBS)	HyClone	SH30088.03
L-glutamine Biowest X0550	Biowest	X0550
Penicillin-Streptomycin	Biowest	L0022
Hygromycin B	Invitrogen	10687-010
C57BL/6J female mice	In Vivos		4-6 wk old
Anesthesia (75 mg/kg Ketamine and 1 mg/kg Medetomidine)	Local pharmacy
Reversal drug (1 mg/kg Atipamezole)	Local pharmacy
Ear punch	Electron Microscopy Sciences	72893-01
Hartmann's solution or Compound sodium lactate	B Braun
Ophthalmic ointment - Duratears sterile ocular lubricant ointment	Alcon
Heat pack - HotHands handwarmers	Heatmax Inc
Introcan Certo IV catheter	B Braun	4251300	24 G x 3/4″
Aquagel Lubricating jelly	Local pharmacy
Poly-L-lysine solution, 0.01%	Sigma	P4707