An orthotopic, postsurgical model of luciferase transfected murine osteosarcoma with spontaneous metastasis

Abstract

Osteosarcoma (OS) is the most common bone tumor in humans. Newer, more clinically relevant models of OS are required to investigate novel therapeutics. The ability to study spontaneous micrometastases in the absence of a primary tumor is important since this is the manner in which most patients are treated clinically. Therefore, we have developed a novel model of murine OS using the DLM8 cell line, which is syngeneic to C3H mice. We have engineered these cells to express firefly luciferase so the development of metastases can be followed serially and non-invasively. These cells form osteolytic/osteoproductive lesions and metastasize spontaneously after orthotopic implantation in the proximal tibia, and the development of soft-tissue metastasis can be followed serially by luciferase expression following amputation. We have demonstrated a significant prolongation of disease-free and overall survival in the surgical adjuvant setting following treatment with doxorubicin or carboplatin, drugs which form the mainstay of treatment for human OS. In conclusion, we have developed a novel surgical adjuvant model of metastatic OS in immunocompetent mice that closely recapitulates the clinical situation, allowing the evaluation of novel therapeutics in the context of minimal residual disease.

Bone Formation Deregulations Are the Oncogenesis Keys in Osteosarcomas

“…All these animal models are outstanding tools to perform in vivo target validation, drug optimization and pre-clinical studies. Furthermore, with the new technologies using bioluminescence, the drug testing and in situ tumor follow up is becoming easier (Sottnik et al, 2010 ;Rousseau et al, 2010).…”

Glycolysis inhibition by 2-deoxy-d-glucose reverts the metastatic phenotype in vitro and in vivo

“…The DLM8-luc-M1 murine OS cells [16] were maintained in C/10 Dulbecco's modified Eagle media [DMEM (Lonza, Walkersville, MD) supplemented with 19 modified Eagle media (MEM) vitamin solution (Cellgro, Henderson, VA), 2 mM L-glutamine (Cellgro), 1 mM sodium pyruvate (Cellgro), 19 non-essential amino acid solution (Cellgro), 19 antibiotic/antimycotic (Cellgro), and 10% heat inactivated FBS (Atlas, Fort Collins, CO)]. Experiments were also performed using C/0.1 media, consisting of 0.1% FBS instead of 10%.…”

Do Mesenchymal Stromal Cells Influence Microscopic Residual or Metastatic Osteosarcoma in a Murine Model?

“…Several rodent models of osteosarcoma have been developed to study pulmonary metastasis [2,6,16,18,19,27]. These models vary widely from scenarios where the primary tumor remains in situ throughout the study to others where the primary tumor is resected before analyzing metastatic growth to models where cell lines are injected into the right ventricle to seed lungs directly.…”

“…These models vary widely from scenarios where the primary tumor remains in situ throughout the study to others where the primary tumor is resected before analyzing metastatic growth to models where cell lines are injected into the right ventricle to seed lungs directly. The difference between models makes it difficult to compare results [6,16,18,19,27]. Orthotopic tumor models with spontaneous metastasis are thought to most accurately recapitulate tumor-stroma interactions in the tumor microenvironment [6,27].…”

“…The difference between models makes it difficult to compare results [6,16,18,19,27]. Orthotopic tumor models with spontaneous metastasis are thought to most accurately recapitulate tumor-stroma interactions in the tumor microenvironment [6,27]. MSCs are known to exist in both the stromal tissues and the circulating blood [15].…”