Murine Bioluminescent Hepatic Tumour Model
July 17th, 2010
This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.
Hello everyone. My name is Simon Regenron. I'm one of the surgical registrars in the car cancer research center here in Ireland.
Today I'll be showing you a surgical technique of inducing liver metastases. In a neuron model, the liver is a common site of metastatic spread and a major cause of cancer related mortality. Only a small percentage of patients are suitable for surgical resection with advanced liver disease.
Therefore, novel therapeutics are needed for the treatment of this condition. Many animal models of hepatic metastases do not result in disease resembling the clinical setting. The model I'm showing you today is a clinically relevant model and it is relatively easy to learn and with very good success rates.
This model involves dividing the spleen into two hemi spleens. One Hemi spleen is used for the inoculation of tumor cells. These tumor cells travel through the porter circulation into the liver.
The other Hemi spleen is left intact maintaining its immunological properties. The cell line I'm using is stably expressing luciferase, allowing for the bioluminescent analysis of tumor growth in vivo. Therefore, this model will allow for the non-invasive monitoring of therapeutics of liver disease.
Okay, let's go ahead and start this for the procedure today. The instruments that we need are dissecting scissors, needle holder, two small faucets, size 10 scalpel five oh proline suture for skin closure, four oh PIA, suture for closure of the peritoneal layer three oh Vicryl ties for dividing the spleen saline 29 gauge insulin needle for anesthetic injection, 26 gauge insulin needle for tumor suspension, injection tumor cell suspension in serum free media, injectable anesthetic, skin marker, precut sterile gauze swabs Ine. The murn model that I will use today is the Athymic MF one nude mouse.
Other models such as the C 57 and BC may be used with appropriate tumor cell lines. A heating pad is used injectable general anesthetic is used in a laboratory. The depth of anesthesia is assessed using the toe pinch.
There should not be a toe withdrawal reflex on toe pinch. The knee size mouse is properly positioned and taped, and abdomen is prepped with Betadine and draped in a sterile fashion. A subcostal incision is made, an abdominal cavity is entered, the spleen is identified and exteriorized.
The spleen is isolated from the rest of the mouse using a precut sterile gauze. Two vertical ties are passed between the two vascular pedicles and gently tied. The spleen is divided into two equal hemi spleens.
One hemi spleen is used for the inoculation of Tumor cells. Tumor cells are slowly injected into the Hemi spleen to avoid leakage. Tumor cells travel through the portal circulation and arrive at the liver.
After 10 Minutes of incubation, the injected hemi spleen is excised and the other hemi spleen is returned into the abdomen. Absorbable sutures and a running stitch are used to close the peritoneal cavity and non-absorbable five oh proline suture is used for skin closure. Recovery is carried out routinely.
The development of liver disease can be assessed over time using 2D ivus imaging. The mouse is anesthetized as before, and Lucifer substrate is given intraperitoneal. The mouse is placed on the stage in the 37 degree chamber.
Imaging is carried out for three minutes and a superimposed image of mouse with detected luminescence is produced Three weeks after treatment, abdomen is opened and diffuse liver disease is found. In conclusion, this technique permits the induction of liver localized metastatic tumor cell growth by luminescent imaging enables visualization and quantification of tumor growth and application in assessment of therapeutic responses.
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Overview
This article describes a surgical technique for inducing orthotopic bioluminescent liver tumors in mice. The procedure allows for the analysis of tumor growth confined to the liver using live whole body luminescence imaging.
Key Study Components
Area of Science
- Oncology
- Neuroscience
- Surgical techniques
Background
- Liver is a common site for metastatic spread in cancer.
- Advanced liver disease limits surgical resection options.
- Understanding liver metastases is crucial for improving patient outcomes.
- Live imaging techniques enhance the study of tumor dynamics.
Purpose of Study
- To demonstrate a method for inducing liver tumors in a mouse model.
- To facilitate the study of tumor growth and behavior.
- To provide a platform for future cancer research.
Methods Used
- Induction of bioluminescent liver tumors in mice.
- Live whole body luminescence imaging for tumor analysis.
- Surgical techniques for tumor induction.
- Monitoring tumor growth over time.
Main Results
- Successful induction of liver tumors in the mouse model.
- Effective monitoring of tumor growth using imaging techniques.
- Insights into the progression of liver metastases.
- Potential applications for therapeutic interventions.
Conclusions
- The surgical technique provides a reliable model for studying liver tumors.
- Live imaging is a valuable tool for monitoring tumor dynamics.
- Further research can build on this model to explore treatment options.
Frequently Asked Questions
Executive Industry Relevance
This model enables biopharma R&D teams to establish orthotopic liver metastases with luciferase-expressing tumor cells, providing a clinically relevant system for evaluating therapeutic interventions. Bioluminescent imaging allows non-invasive, longitudinal monitoring of tumor burden, supporting mechanistic de-risking and predictive confidence in lead identification. The surgical approach ensures reproducible tumor engraftment via portal circulation, facilitating standardized preclinical assessment of liver metastasis therapeutics.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Scientific Value: Enables interrogation of metastatic tumor-cell survival and growth mechanisms in a liver-specific microenvironment.
- Operational Value: Provides a reproducible surgical model for consistent tumor induction across study cohorts.
Screening & Assay Development
- Scientific Value: Generates quantifiable bioluminescent signals correlating with viable tumor cell burden for dose-response assessment.
- Operational Value: Supports standardized imaging protocols using IVIS for longitudinal therapeutic efficacy screening.
Translational & Preclinical Research
- Scientific Value: Models hepatic metastasis progression relevant to human disease, enabling evaluation of tumor microenvironment interactions.
- Operational Value: Allows early detection of treatment-induced tumor necrosis via bioluminescent signal reduction, informing go/no-go decisions.
Pipeline & Workflow Integration
The model integrates into the discovery continuum from target validation through preclinical efficacy testing, supporting liver metastasis therapeutic development.
- Discovery Biology: Supports hypothesis testing of metastatic mechanisms using luciferase-labeled tumor cells in an orthotopic liver context.
- Screening: Enables assay-ready tumor models with standardized bioluminescent readouts for compound library evaluation.
- Analytics: Provides quantitative tumor growth measurements via IVIS imaging, facilitating statistical comparison of treatment groups.
- Translational Research: Connects discovery findings to preclinical validation through clinically relevant liver metastasis phenotypes.
- Enterprise Reuse: Establishes a reusable surgical and imaging platform for multiple oncology programs targeting hepatic metastases.
Operational & Enterprise Impact
- Scientific Value: Mechanistic de-risking of liver metastasis hypotheses through direct visualization of tumor growth and treatment response.
- Operational Value: Standardized surgical procedure and imaging workflow ensure reproducibility across sites and studies.
- Strategic Value: Enables earlier, data-driven go/no-go decisions by quantifying therapeutic impact on tumor burden.
- Portfolio Impact: Supports risk-adjusted prioritization of liver metastasis candidates based on predictive bioluminescent endpoints.
Implementation Considerations
- Requires expertise in murine surgical techniques, including spleen mobilization and hemi-spleen resection.
- Dependent on IVIS imaging infrastructure and luciferase substrate administration for bioluminescent detection.
- Necessitates standardization of tumor cell inoculation volume and incubation timing across experimental groups.
- Requires aseptic surgical practices to maintain model integrity and reduce variability.
- Limited to immunocompromised mouse models (e.g., athymic nude) for human tumor cell engraftment, which may affect immune-related therapeutic evaluations.
