Related Experiment Video
Updated: Dec 25, 2025
A Murine Orthotopic Bladder Tumor Model and Tumor Detection System
Published on: January 12, 2017
Bilateral murine tumor models for characterizing the response to immune checkpoint blockade
Rachael M Zemek1,2,3, Vanessa S Fear4,5,6, Cath Forbes4,5,6
1School of Biomedical Sciences, University of Western Australia, Crawley, Western Australia, Australia. rachael.zemek@telethonkids.org.au.
Abstract:
The therapeutic response to immune checkpoint blockade (ICB) is highly variable, not only between different cancers but also between patients with the same cancer type. The biological mechanisms underlying these differences in response are incompletely understood. Identifying correlates in patient tumor samples is challenging because of genetic and environmental variability. Murine studies usually compare different tumor models or treatments, introducing potential confounding variables. This protocol describes bilateral murine tumor models, derived from syngeneic cancer cell lines, that display a symmetrical yet dichotomous response to ICB. These models enable detailed analysis of whole tumors in a highly homogeneous background, combined with knowledge of the therapeutic outcome within a few weeks, and could potentially be used for mechanistic studies using other (immuno-)therapies. We discuss key considerations and describe how to use two cell lines as fully optimized models. We discuss experimental details, including proper inoculation technique to achieve symmetry and one-sided surgical tumor removal, which takes only 5 min per mouse. Furthermore, we outline the preparation of bulk tissue or single-cell suspensions for downstream analyses such as bulk RNA-seq, immunohistochemistry, single-cell RNA-seq and flow cytometry.
Insights
New mouse models show symmetrical yet dichotomous responses to immune checkpoint blockade (ICB) therapy. These models aid in understanding the variable therapeutic response to ICB by enabling detailed analysis in homogeneous backgrounds.
Area of Science:
- Immunology
- Oncology
- Translational Research
Background:
- Therapeutic response to immune checkpoint blockade (ICB) varies significantly across cancer types and patients.
- Biological mechanisms driving differential ICB response remain poorly understood.
- Existing murine models present challenges due to confounding variables from genetic and environmental heterogeneity.
Purpose of the Study:
- To develop and present a protocol for bilateral murine tumor models with symmetrical yet dichotomous responses to ICB.
- To enable detailed analysis of tumors within a homogeneous genetic and environmental background.
- To facilitate mechanistic studies for optimizing immunotherapy strategies.
Main Methods:
- Utilized bilateral murine tumor models derived from syngeneic cancer cell lines.
- Employed precise inoculation techniques for symmetrical tumor growth.
- Performed one-sided surgical tumor removal for comparative analysis.
- Prepared bulk tissue and single-cell suspensions for downstream molecular analyses.
Main Results:
- Established optimized models using two specific cell lines demonstrating predictable, dichotomous ICB responses.
- Achieved symmetrical tumor growth and response patterns within the bilateral model.
- Successfully generated tissue and single-cell suspensions suitable for various analyses.
Conclusions:
- The described bilateral murine tumor models offer a robust platform for investigating ICB response variability.
- These models facilitate in-depth mechanistic studies by minimizing confounding factors.
- The protocol supports diverse downstream analyses, including transcriptomics and flow cytometry, advancing immunotherapy research.
