A validated mouse model for orthotopic bladder cancer using transurethral tumour inoculation and bioluminescence imaging

Abstract

OBJECTIVES To describe a technique for transurethral tumour inoculation, bioluminescence imaging (BLI) and validation of this approach using ex vivo magnetic resonance imaging (MRI), as a reproducible and quantifiable model of orthotopic bladder cancer is required to enable preclinical pharmacological studies of intravesically administered anticancer agents and the use of BLI provides a sensitive method to monitor tumour growth over time. MATERIALS AND METHODS Human KU‐7 bladder tumour cells were transduced with a lentiviral construct to stably express the firefly luciferase gene. These cells were then inoculated in female nude mice by intravesical instillation. BLI was performed weekly and the mice were killed after 4 weeks. Ex vivo MRI and whole‐mount step‐sections were obtained to assess bladder tumour volume. RESULTS KU‐7 tumour cells were highly tumorigenic and were successfully inoculated in 96% of mice. After 4 weeks, all tumours were confined to the mucosa and submucosa (≤pT1). There was an excellent correlation between tumour volume and BLI for both ex vivo bladder MRI (R2 = 0.929) and end‐point histological measurements (R2 = 0.836). CONCLUSIONS We have established and validated a reliable model of orthotopic bladder cancer that can be used to evaluate various methods of intravesical therapy. BLI allows excellent longitudinal surveillance and quantification of tumour burden.

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

“…For in vivo growth analysis under different treatment strategies murine orthotopic bladder cancer models remain the reference standard 1,2 . The inoculation of human bladder cancer cells in immunocompromised mice (xenograft model) relies on intravesical instillation ("intravesical model") 3,4,5 or direct injection into the bladder wall ("intramural model") 6,7 . Both techniques can also be performed in rats 8,9 .…”

Intravesical α-Radioimmunotherapy with 213Bi-Anti-EGFR-mAb Defeats Human Bladder Carcinoma in Xenografted Nude Mice

“…Because we could not detect any abdominal spread of tumor cells via bioluminescence imaging, bladder perforation due to cauterization could be excluded at any time. Abdominal tumor spread has been observed in another orthotopic xenograft model not using cauterization (8).…”

“…To monitor tumor growth and therapeutic efficacy, noninvasive imaging concepts are preferable. For that purpose, tumor cells are stably transfected with genes coding for fluorescent proteins (7) or enzymes catalyzing bioluminescence (8), allowing for the continuous visualization of tumor development after intravesical instillation of tumor cells.…”

Animal Models for Basic and Preclinical Research in Bladder Cancer

“…Also, analysis of the response to cytotoxicity and immunotherapy treatments can be performed (Bhattacharya et al, 2010;Takeuchi et al, 2011;Zhang et al, 2011). cells are stably transfected with genes coding for fluorescent proteins (Tanaka et al, 2003) or enzymes catalyzing bioluminescence (Hadaschik et al, 2007), allowing for the continuous visualization of tumor development after intravesical instillation of tumor cells. Although, as described above, xenograft models are important tools to study the behavior of human tumors in vivo, they also have an important limitation: they are immunodeficient.…”