To begin, take the freshly resected osteosarcoma tissue, stored in the tissue protective solution. Rinse the osteosarcoma tissues twice with pre-cool sterile physiological saline in a sterile hood. Then remove the areas of osteosarcoma tissue with bleeding and necrosis.
Using a scalpel, cut the clean osteosarcoma tissue in a tissue culture dish containing pre-cool sterile physiological saline. Keep the tissue on ice. Now, place the nude mice on sterile surgical drapes.
Pinch the toes of the mice to ensure that they are completely anesthetized. Then secure the mouse in a lateral decubitus position. With a marker pen, mark the surgical incision site on the skin.
Use the scalpel to make a five millimeter incision from the skin to the subcutaneous tissues. Now, hold ophthalmic tweezers in the left hand and lift the upper margin of the incision. Simultaneously, use a straight needle holder with the right hand to perform blunt dissection upward under the dermis of the mice.
With ophthalmic tweezers, hold the upper side skin of the incision. Then transplant the osteosarcoma tissue under the skin five millimeters above the incision margin. Suture the incision using surgical lines, with two to three stitches for a five millimeter incision.
Transplanting femoral osteosarcoma tissue from a 15-year-old patient to P0, P1, P2, and P3 mice resulted in the successful creation of a patient-derived xenograft.
