Bioluminescent Imaging of Breast Tumor Progression in a Female Mouse Model

Abstract

Source:
Marina Arantes Radicchi¹, Victor Carlos Mello¹,
Victória Paz Machado¹, Márcia Cristina Oliveira da Rocha¹, Jaqueline Vaz de Oliveira¹, Luana Cristina Camargo^1,2, Luís Alexandre Muehlmann¹, Ricardo Bentes Azevedo¹, Sônia Nair Bao¹, João Paulo Figueiró Longo<sup>1

1</sup>Institute of Biological Sciences, University of Brasília, ²Department of Basic Psychological Processes, Institute of Psychology, University of Brasília

This video demonstrates the procedure for tracking tumor progression in vivo by injecting luciferase-expressing breast adenocarcinoma cells into a female mouse and monitoring bioluminescent signals to evaluate primary and metastatic tumor growth over time

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Cell culture

1. Maintain 4T1 cell line (ATCC-CRL-2539), a murine breast adenocarcinoma cell, in culture with DMEM medium (Dulbecco's modified eagle medium) supplemented with 10% fetal bovine serum (v:v), penicillin (100 U/mL), and streptomycin (100 U/mL), at 37 °C and in a humidified atmosphere containing 5% CO₂.
2. Transfect the original cell line with the Luc2 gene, which expresses luciferase. This bioluminescent enzyme is constitutively produced.

2. Animal model - Tumor....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
D-luciferin	Perkin Elmer	122799	Chemical Reagent
Dulbecos’s modified Eagle’s médium	Gibko	A7030	Cell Culture Media
Fetal Bovine Serum	Merck	F2442	Cell Culture Media Supplement
Isoflurane	Cristália	NA	Anesthetics
IVIS Lumina	XR	Series III	Equipment – USA
Penicillin	Merck	P4333	Cell Culture Antibiotic
Phalloidin-Alexa Fluor 488	Thermo Fischer	A12379	Chemical Reagent
Phosphate buffer saline	Merck	4437568	Chemical Reagent
Streptomycin	Merck	P4333	Cell Culture Antibiotic