Delivering Effector T Lymphocytes into Mice Carrying Human Brain Tumors

Take an anesthetized immunodeficient mouse carrying an engrafted human brain tumor.

Remove the fur from the surgical site and disinfect. Apply an ointment on the eyes to prevent corneal drying.

Position the mouse on a stereotactic frame with a heating pad to maintain body temperature.

Make an incision on the scalp, exposing the skull.

Locate the bregma, a landmark on the skull, and determine the injection location to position a microsyringe above it.

The microsyringe contains a suspension of tumor-specific effector T lymphocytes from healthy human donors.

Create a hole at the injection location on the skull, avoiding brain injury. Inject the needle into the brain and slowly release the effector T lymphocytes at the tumor site.

Partially withdraw the needle and keep it in place, preventing the leakage of injected cells, before withdrawing the needle completely.

Close the incision and allow the mouse to recover.

Ensure that the animal is adequately anesthetized by a toe pinch, and then remove fur from the surgical site between the two ears and up to the nose. Resuspend cells slowly to prevent cell clumping, and load cell suspension into syringe with great care to avoid the presence o