In vitro and in vivo experimental models of canine osteosarcoma have been described.1–11 In vivo models that use mice are superior to in vitro models in that tumor characteristics more closely approximate spontaneously developing disease. Models that use mice also facilitate rapid advances in research by generating large numbers of reproducible neoplasms in a short period at a low financial cost, allowing large volumes of data to be gathered without the need for dogs with osteosarcoma.2, 12 Many methods of experimentally inducing canine osteosarcoma in mice have been reported and involve SC injection in the interscapular region, proximal aspect of the hind limb, and abdominal or flank region as well as orthothotopic femoral xenotransplantation.2–11 However, these models have various inadequacies, including development of irregular tumor size and shape, adverse local xenograft sequelae (eg, ulceration or hemorrhage), and difficulty of antemortem tumor measurement.2–11
External beam radiation therapy is used as a palliative and curative treatment in dogs with appendicular osteosarcoma; however, to our knowledge, an in vivo radiation model for canine osteosarcoma has not been developed. To reduce radiation-associated adverse effects associated with experimental in vivo induction of osteosarcoma via xenografts, distal appendicular tumors are desirable for such a model.13–16 Similar IM xenografts have been developed to simulate human tumors, including multiple idiopathic hemorrhagic sarcoma (Kaposi's sarcoma), breast cancer, and melanoma.13–16
The purposes of the study reported here were to establish a reproducible, distal appendicular, IM xeno-graft model of canine osteosarcoma in mice; to characterize the morphology of the tumors induced; to establish tumor growth curves for specific doses of radiation; and to determine percentages of tumor necrosis that result from specific doses of radiation. Our hypotheses were that 1 radiation dose of 10 or 15 Gy would significantly delay tumor growth and that radiation therapy would induce dose-dependent tumor necrosis. The usefulness of such a model is that it could be used to evaluate the effectiveness of various radiation therapy protocols and the effects of radiopotentiating Agents. Furthermore, the IM xenograft model could serve as a translational model for human osteosarcoma, given the biologic and histologic similarities between canine and human osteosarcoma.17
Highly metastasizing parent osteosarcoma
BALB/c AnNCrl-nuBR mice, Charles River Laboratories, Wilmington, Mass.
Cells provided by Dr. Tsuyoshi Kadosawa, Laboratory of Veterinary Surgery, Hokkaido University, Sapporo, Japan.
RPMI 1640 without L-glutamine, Mediatech Inc, Herndon, Va.
Regular (heat-inactivated) fetal bovine serum, Mediatech Inc, Herndon, Va.
Penicillin-streptomycin solution (100X), Mediatech Inc, Herndon, Va.
Liquid L-glutamine, Mediatech Inc, Herndon, Va.
MEM vitamins, Mediatech Inc, Herndon, Va.
MEM nonessential amino acids, Mediatech Inc, Herndon, Va.
Nunc EasYFlasks Nunclon, Thermo Fisher Scientific, Roskilde, Denmark.
Hanks' balanced salt solution without calcium and magnesium, Mediatech Inc, Herndon, Va.
Mediatech Inc, Herndon, Va.
PBS solution without calcium and magnesium, Mediatech Inc, Herndon, Va.
R-2040 Template Metric Circle, Berol Rapidesign, Leeds, Mass.
Varian Oncology Systems, Palo Alto, Calif.
Super Up-Rite Bond-Rite microscope slides, Richard-Allan Scientific, Kalamazoo, Mich.
Richard-Allan Scientific, Kalamazoo, Mich.
Princess pressure cooker, Cell Marque Corp, Rocklin, Calif.
Lab Vision, Freemont, Calif.
UltraVision Detection System anti-rabbit HRP/DAB, Lab Vision, Freemont, Calif.
Haematoxylin 7211, Richard-Allan Scientific, Kalamazoo, Mich.
Clarifier 2, Richard-Allan Scientific, Kalamazoo, Mich.
Richard-Allan Scientific, Kalamazoo, Mich.
Histology Tech Services Inc, Gainesville, Fla.
Olympus BX41 microscope with attached Olympus DP70 10 megapixel camera, Olympus Optical Co Ltd, Center Valley, Pa.
DPController, version 126.96.36.199 and DPManager, version 188.8.131.52, Olympus Optical Co Ltd, Center Valley, Pa.
UTHSCSA Image Tool for Windows, version 3.00, University of Texas Health Science Center, San Antonio, Tex.
SigmaStat for Windows, version 3.0, SPSS Inc, Chicago, Ill.
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