Honors College Thesis
 

Visualizing Internalization of Mesenchymal Stem Cell-Derived Extracellular Vesicles by Canine Osteosarcoma Cells

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  • Osteosarcoma (OSA) is a primary malignant bone tumor sarcoma that occurs in dogs as well as people and has aggressive biologic behavior with devastating consequences. Because of the similarities in morphology and prognosis, OSA in dogs represents an effective model for the equivalent disease in humans. Mesenchymal stem cells (MSCs) are multipotent adult stem cells that can be harvested from fat and bone marrow. Local tissue MSCs are vital for homeostasis and are recruited from more distant tissues and organs to tumor sites. MSCs have potential for the mass production of extracellular vesicles (MSC-EVs) for use in anti-tumor drug delivery. An initial step in establishing the therapeutic use of MSC-EVs for treatment of OSA is confirmation that OSA tumor cells uptake MSC-EVs. The objective of this study was to visualize MSC-EV internalization by OSA cells, with the hypothesis that internalization occurs within three hours of exposure. OSA cells were obtained from canine patients with primary bone tumors undergoing routine amputation. MSCs were obtained from discarded adipose tissue during routine ovariohysterectomy in healthy dogs. MSC-EVs were isolated, labeled with a fluorescent dye, washed, and plated on coverslip monolayers of OSA cells. After a three-hour incubation, coverslips were labeled with a nucleic-acid fluorescent dye and visualized using fluorescence microscopy. Fluorescence was identified in the cytoplasm of OSA cells treated with MSC-EVs, while negative-control OSA cells lacked evidence of non-specific fluorescence. These findings confirm the hypothesis that MSC-EV uptake occurs within three hours and can be visualized for confirmation.
  • Key Words: cancer, dog, mesenchymal stem cell, osteosarcoma, extracellular vesicle
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