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. 2025 Jan 11;17(1):e77291.
doi: 10.7759/cureus.77291. eCollection 2025 Jan.

Standardization of a Model of Vertebral Metastasis of Breast Cancer in CD1/Nu/Nu Mice

Gervith Reyes Soto  1 , Vladimir Miranda-Galván  2 , Norma Uribe-Uribe  3 , Juan Manuel Escobar-Valderrama  2 , Jorge Alanis-Mendizabal  2 , Luis A Medina-Velázquez  4 , Alejandro Garcia  5 , Gonzalo Torres Villalobos  2 , Fabian Díaz-Martínez  6 , Paola Montiel de la Rosa  2 , Carlos Bravo-Reyna  2 , Alejandra Guadalupe Cervantes Zentella  2 , Geovanny Jose Vanegas Cerna  7 , Vladimir Nikolenko  8 , Tshiunza Cherubin  9 , Andreina Rosario Rosario  10 , Carlos Castillo-Rangel  11 , Mario Antonio Furcal Aybar  12 , Laith Wisam Alsaed  13 , Manuel De Jesus Encarnacion Ramirez  13   14
Affiliations

Standardization of a Model of Vertebral Metastasis of Breast Cancer in CD1/Nu/Nu Mice

Gervith Reyes Soto et al. Cureus. .

Abstract

Introduction: Breast cancer is the leading cause of cancer-related death in Mexico, with high mortality associated with spinal bone metastasis. We propose to standardize a murine model of bone metastasis to study and understand the tumor microenvironment.

Materials and methods: An experimental, prospective, longitudinal study was conducted using 18 CD1/Nu/Nu 30g nude mice. Two cell lines, MCF-7 and 4T1, were inoculated, clinical follow-up was performed, and biopsy samples were obtained for histopathological evaluation.

Results: Histopathological evaluation of models inoculated with the MCF-7 cell line showed no tumor development, while inoculation with the 4T1 cell line resulted in tumor development, as evidenced by PET-CT and histopathology, using 5,000 and 1,000 cells, respectively.

Conclusions: The use of this model is proposed for studying the clinical, molecular, and prognostic aspects of breast cancer progression by inoculating 1,000 cells of the 4T1 cell line.

Keywords: 4t1; breast cancer; hematogenous metastasis; pain; pet-ct; spinal cord compression.

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Conflict of interest statement

Human subjects: All authors have confirmed that this study did not involve human participants or tissue. Animal subjects: Bioethics Committee of National Cancer Institute, Mexico Issued protocol number NOM 062-ZOO-1999. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. Surgical approach: A) evisceration and identification of the psoas and vascular structures; B) dissection of the psoas muscle and localization of the lumbar vertebral body; C) perforated lumbar vertebral body, exposed for inoculation of cancer cells.
Figure 2
Figure 2. Histopathological sections with H&E staining of the spinal column of mice inoculated with breast cancer cells from the human MCF-7 cell line, showing cells with a necrotic appearance in the bone marrow, without identifying tumor-origin cells. A) Mouse 1; B) Mouse 2; C) Mouse 3.
Figure 3
Figure 3. PET-CT performed in the model three weeks post-surgery, showing evidence of 18-FDG uptake as the contrast agent in the lumbar vertebrae at the site where the tumor cells were inoculated, along with areas of external uptake, which are attributed to a probable infectious process.
Figure 4
Figure 4. Macroscopic samples obtained after sacrificing the experimental models upon reaching the humanitarian endpoint, showing tumor development in the lumbar vertebrae with invasion into adjacent organs.
Figure 5
Figure 5. Histopathological sections with H&E staining of the spine of mice inoculated with murine 4T1 breast cancer cells. A) Presence of tumor cells in tissues adjacent to the spine (4x); B) tumor cells in pancreatic tissue, metastatic to the spine (10x); C) tumor cells in the vertebral bodies (4x); D) enlargement of lamella C, showing tumor cells in the striated muscle of the paravertebral musculature (10x).
See this image and copyright information in PMC

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