J Korean Soc Radiol.  2011 Jan;64(1):57-65. 10.3348/jksr.2011.64.1.57.

A Bone Metastasis Nude Mouse Model Created by Ultrasound Guided Intracardiac Injection of Breast Cancer Cells: the Micro-CT, MRI and Bioluminescence Imaging Analysis

• Park YJ
• Song EH
• Kim SH
• Choi SH
• Song HT
• Suh JS

• KMID: 1443583
• DOI: http://doi.org/10.3348/jksr.2011.64.1.57

Abstract

PURPOSE
The purpose of this study was to develop a nude mouse model of bone metastasis by performing intracardiac injection of breast cancer cells under ultrasonography guidance and we wanted to evaluate the development and the distribution of metastasis in vivo using micro-CT, MRI and bioluminescence imaging.
MATERIALS AND METHODS
Animal experiments were performed in 6-week-old female nude mice. The animals underwent left ventricular injection of 2x105 MDA-MB-231Bo-Luc cells. After injection of the tumor cells, serial bioluminescence imaging was performed for 7 weeks. The findings of micro-CT, MRI and the histology were correlated with the 'hot' lesions seen on the bioluminescence imaging.
RESULTS
Metastasis was found in 62.3% of the animals. Two weeks after intracardiac injection, metastasis to the brain, spine and femur was detected with bioluminescence imaging with an increasing intensity by week 7. Micro-CT scan confirmed multiple osteolytic lesions at the femur, spine and skull. MRI and the histology were able to show metastasis in the brain and extraskeletal metastasis around the femur.
CONCLUSION
The intracardiac injection of cancer cells under ultrasonography guidance is a safe and highly reproducible method to produce bone metastasis in nude mice. This bone metastasis nude mouse model will be useful to study the mechanism of bone metastasis and to validate new therapeutics.

MeSH Terms

Animal Experimentation
Animals
Brain
Breast
Breast Neoplasms
Disease Models, Animal
Female
Femur
Humans
Luminescent Measurements
Magnetic Resonance Imaging
Mice
Mice, Nude
Neoplasm Metastasis
Skull
Spine

Figure

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  Fig. 1 Ultrasonogram of intracardiac injection to nude mouse. Ultrasound guided intracardiac injection to the left ventricle of nude mouse is shown. The needle tip is seen inside of the left ventricle.

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  Fig. 2 Bioluminescence photon intensity of serially diluted MDA-MB-231Bo-Luc cell line in 96-well plate. Cells were serially diluted in from 1×105 to 7.8×103 cells/well. Wells with media (no cells) and distilled water (D.W.) were included as control.

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  Fig. 3 Growth kinetics of MDA-MB-231Bo-luc cell line in individual mice. A. Bioluminescence images from a representative mice, taken from the dorsal side, with every week intervals after injection of 2 × 105 MDA-MB-231Bo-Luc cells. Brain, femur and spine metastasis was detected. B. Serial average photon flux intensity from all animal.

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  Fig. 4 Multimodal imaging showing femur metastasis. 3D (left) and 2D (right small figures in prone, supine, right lateral, and left lateral positions) bioluminescence images, signal was shown on brain and right femur. Micro-CT image shows cortical osteolysis of the right femur (arrows). The 3D reconstructed image of micro-CT shows lesion extent of bone metastasis on the femur (arrows). Axial T2 weighted MR image shows extraskeletal extension of the metastatic tumor (arrow).

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  Fig. 5 Multimodal imaging showing spine and brain metastasis. 3D bioluminescence image shows brain and spine metastasis. Micro-CT image shows osteolysis at the left temporal bone (arrows). Micro-CT image of the spine shows cortical destruction on the spine (arrow). Sagittal T2 weighted MR image shows high signal intensity intracranial and spinal canal mass lesions (arrows).

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  Fig. 6 Histopathology of the bone and brain. Hematoxylin-eosin staining of brain (A, B), femur (D, E), and spine (G, H) and corresponding immunohistochemical staining(C, F, I), respectively. Insets in A, D, G are magnified in B, E, H. Magnification is × 40 (A, D, G) or × 200 (B, C, E, F, H, I).

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