This article details the procedure for intracranial injection of cancer cells in mice, a method used to monitor tumor growth in the brain. The process involves precise surgical techniques to ensure accurate cell delivery while minimizing tissue damage.
Intracranial injection of cancer cells is a procedure to inject cancer cells into the brain through the cranium, the bone that protects the brain. This procedure helps monitor tumor growth in the mouse brain.
To begin, position an anesthetized mouse with a calvarial window - surgically created burr hole in the skull - onto a stereotactic unit.
Next, prime a syringe containing homogenous cell suspension to remove air bubbles.
Wipe the outer barrel of the needle shaft with an alcohol swab to prevent tumor cell seeding along the injection tract.
Now, load the syringe on the automatic injector apparatus attached to the stereotactic unit.
Align the needle to the center of the calvarial window close to the exposed cerebrum - the uppermost portion of the brain.
Insert the needle briefly into the brain and hold it there to acclimatize the brain tissues around it.
Slowly inject the cells into the brain to prevent any tissue damage and let the needle rest in the organ to restrict the backflow of tumor cells.
Thereafter, carefully remove the needle to prevent tumor cells from tracking up the needle tract.
Apply bone wax to the calvarial window to retain the tumor within the brain.
Last, seal the incision with suture clips and allow the mouse to recover in a warm cage.
For cancer cell injection, attach the automatic injector unit to the stereotactic apparatus and load 6 to 8 microliters of cells, thoroughly resuspended in DPBS, into a sterile Hamilton syringe.
Load the syringe onto the injector and dispense a small volume of solution onto a disposable sterile drape to prime the needle for injection.
Use a cotton tip applicator to wipe the syringe with 70% ethanol and align the needle tip to the center of the calvarial window until it nearly touches the exposed cerebrum.
Reset the digital Vernier scale to zero and slowly insert the needle into a 3-millimeter depth into the brain. Leave the needle in the brain for at least 60 seconds before selecting RUN on the injector screen to begin the cell delivery to the injection site.
When all of the cells have been delivered, allow the needle to rest in the brain for at least 3 minutes to allow the brain parenchyma to acclimate to the injection, before retracting the needle from the brain at a rate of 0.75% millimeters per minute.
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