Establishing a Murine Superficial Bladder Cancer Model via an Intravesical Cell Administration Technique

Summary

This protocol describes a unique method for constructing an orthotopic model of superficial bladder cancer.

Abstract

This study presents an innovative method for establishing an orthotopic murine bladder tumor model with high efficiency and precise tumor localization. After anesthetizing female C57BL/6J mice in a supine position, a 24 G intravenous needle is inserted into the bladder to evacuate its contents. A 34 G dispensing needle is then introduced through the catheter, rotated five times to create a focal injury to the bladder dome mucosa, and subsequently removed. The MB49 cell suspension is aspirated and connected to a 30 G dispensing needle, which is inserted into the bladder via the catheter. Tumor cells are injected submucosally into the bladder under pressure. This technique results in minimal trauma to the mice, a high tumor take rate, and a fixed tumor location. It is characterized by simplicity and excellent reproducibility. This model provides an ideal experimental platform for developing intravesical therapies for bladder cancer, facilitating the advancement and optimization of treatment strategies for this malignancy.

Introduction

Bladder cancer represents a significant global health burden, with notable sex-specific disparities in incidence and prognosis. This malignancy is characterized by distinct molecular subtypes, each associated with diverse pathogenic pathways. The molecular and pathological features differ significantly between non-muscle invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC)^1,2. NMIBC accounts for approximately 75% of cases, featuring tumors of transitional epithelial origin that remain localized without metastasis or spread. Conversely, MIBC is characterized by the infiltration of cancer ce....

Protocol

All animal experimental protocols and procedures were approved by the Ethical Review Committee for Animal Experimentation of Tianjin Medical University, Tianjin, China (approval number SYXK: 2020-0010). Six- to eight-week-old female C57BL/6J mice were used for this study. The animals were housed under controlled environmental conditions, including a 12-h light-dark cycle, temperatures ranging from 21-25 °C, adjustable humidity levels between 30%-70%, and unrestricted access to food and water unless otherwise specifi.......

Discussion

Research on bladder cancer relies on animal models, which are indispensable for both basic and applied research. To better mimic the tumor growth environment, orthotopic bladder tumor models provide a superior approach compared to subcutaneous tumor models¹¹.

Currently, orthotopic bladder cancer models can be categorized based on experimental purposes into two main types: those utilizing immunodeficient mice, such as patient-derived xenograft (PDX) models

Materials

Name	Company	Catalog Number	Comments
34 G dispensing needle	Suzhou Haorun Fluid Technology Co., Ltd., Suzhou, China	SGL-362
30 G dispensing needle	Suzhou Haorun Fluid Technology Co., Ltd., Suzhou, China	SGL-362
Avertin	Sigma-Aldrich LLC	T48402
Trypan blue	Sigma-Aldrich LLC	302643