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Orthotopic inoculation helps deliver cancer cells to a recipient animal at a specific tissue or site matching the cells' natural anatomical location. To begin, prep an anesthetized female nude mouse, belly side down, on an operating stage. Make a vertical incision near its spine at a position between the rib cage and the femur. Open the abdominal muscles to expose the peritoneal cavity partially. Locate one of the ovaries.
Prepare a suspension of ovarian cancer cells with chilled extracellular matrix in a syringe. The low temperature prevents the suspension from solidifying. Inject this suspension into the mouse's ovary. Hold the needle in place to allow the matrix to solidify and embed the cancer cells within the ovary. Retract the needle. Suture the incision to close the wound. Allow the mouse to recover.
Over time, the injected ovarian cancer cells proliferate to form a primary tumor. Eventually, a few cells detach from their primary location and disseminate within the peritoneal cavity. When these migrating tumor cells attach to the peritoneal cavity lining, they invade the peritoneal tissue and proliferate to form metastasized secondary tumors.
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