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

During orthotopic intramucosal inoculation, tumor cells matching the mucosal tissue histotype get delivered into the mucosal tissue of a recipient animal.

To begin, restrain an anesthetized mouse by its neck on a surgical platform for easier access to its mouth.

Prepare a suspension of highly metastasizing cancer cells in a chilled basement membrane matrix in a syringe. ​The low temperature prevents the suspension from solidifying.

Carefully inject this suspension into the inner cheek or buccal region of the mouse. The buccal region is lined by the buccal mucosa, which is surrounded by a network of blood and lymphatic vessels.

Ensure complete delivery of the cancer cell suspension and gently retract the needle. Allow the matrix to solidify and embed the cells within.

Allow the mouse to recover. Within days of injection, a primary tumor develops at the site of injection. Eventually, the cells from this tumor with high metastatic potential may travel through the lymphatic system and extravasate into lymph nodes.

These cancer cells adapt to their new microenvironment and proliferate and establish metastatic colonies in lymph nodes.