Murine Bioluminescent Hepatic Tumour Model

Summary

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.

Abstract

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis.

Protocol

Introduction

The liver is often the first site of metastatic disease and may be the only site of spread in as many as 30 40% of patients with advanced disease. However options for treating metastatic liver tumours are few with a minority of patients suitable for resection and only 23% major responders to chemotherapeutics. Preclinical research in this field has been limited by the absence of a suitable mouse model to study isolated liver disease. We describe a split-spleen approach that can be used to reliably establish uniform, diffuse liver metastases. Resection of this half of the spleen avoids the consequences of splenic tumour growth an....

Discussion

The development of reliable in vivo models for the study of liver metastasis is becoming increasing important with the advent of new therapeutic strategies. For the accurate study of new strategies, animal models should be relatively easy to perform, form a significant number of metastasis, involve all the steps of the metastatic cascade and have a reporter system that allows detection and counting of a limited number of metastatic cells. The split-spleen approach as described involves a single operation and is .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
DMEM	Sigma-Aldrich	D6429
PBS	Sigma-Aldrich	D8537
Ketamine	Vétoquinol
Xylazine	Vétoquinol
Carprofen	Vétoquinol
Videne	Ecolab
Vicryl 4-0 suture	Ethicon Inc.
PDS 4-0 suture	Ethicon Inc.
Prolene 4-0 suture	Ethicon Inc.
Firefly Luciferin	Biosynth International, Inc