Intramucosal Inoculation of Cancer Cells in Murine Model: A Procedure to Establish an Orthotopic Head and Neck Cancer Mouse Model

Immediately before the injection, mix the cells at a 1 to 1 tumor cell suspension to basement membrane matrix ratio on ice. Load one 0.5 milliliter syringe equipped with a 23G needle with 100 microliters of cells per recipient animal. Place the syringes on ice, and confirm a lack of response to toe pinch in an anesthetized mouse.

Next, insert the needle into the right or left buccal region through the available open space on either side of the mouth, keeping the syringe parallel to the buccal region while it is inside the oral cavity, to facilitate injection of the full 100-microliter volume of the cell basement membrane matrix suspension over a period of 5 seconds. Keep the syringe in place for an additional 5 seconds to ensure that all of the material has been injected before withdrawing the syringe gently. The tumors will become grossly visible in about a week.