How to Make an Orthotopic Model of Brain Tumor in Mouse?
(Intracerebral injection protocol for an orthotopic brain cancer model in mouse)
Materials:
1. Surgical scissors, forceps, and scalpel
2. 70% ethanol wipes and Betadine swabs
3. An autoclip wound applier and stainless-steel autoclips
4. Q-tips
5. 27- or 30-gauge needles
6. A 0.3 cc insulin syringe
7. Tumor cells
Anesthetics:
Nembutal (Pentobarbital sodium)
Methods:
1. Anesthetize a mouse by i.p. injection of pentobarbital sodium (50mg/kg body wt).
2. Restrain the mouse on the stomach on a plastic board.
3. Disinfect the skin of the head top with Betadine and 70% ethanol.
4. Incise the skin along the midline for about 0.5 cm to expose coronal and sagittal sutures.
5. At a point on the right site 3mm dorsal to the coronal suture and 3mm lateral to the sagittal suture, insert 28-Gauge needle on a 0.3 cc insulin syringe (Bacton Dikinson Co., Rutheford, NJ) into the right cerebral hemisphere to a depth of 3mm.
6. Inject 10 µl of tumor cell suspension slowly and hold the needle for 30 seconds after injection.
7. Retract the needle.
8. Place a Q-tip over the injection site for about 1 min to prevent bleeding and spillage of material.
9. Close the skin with wound autoclips.
10. Keep the mouse warm during recovery.
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