Intracranial Orthotopic Model: A Procedure to Implant Cancer Cells in Mouse Brain

Abstract

Source: Geisler, J. A. et al. Modeling Brain Metastases Through Intracranial Injection and Magnetic Resonance Imaging. J. Vis. Exp. (2020)

This video demonstrates implantation of tumor cells in the mouse brain by performing intracranial injection.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Injecting Cancer Cells into the Brain Place an anesthetized mouse with an exposed calvarial window on the stereotatic frame. Attach an automatic injector unit to the stereotactic apparatus (Figure 1). Pull up >2 µL of cells thoroughly resuspended in 1x DPBS in a clean Hamilt…

Representative Results

Figure 1: Stereotactic and anesthesia systems. Stereotactic stand with automated syringe pump and mouse elevating box. The box is an inverted pipette tip box containing hand warmers used to elevate the mouse to the appropriate height and maintain body temperature. Inset shows the orientation of the Hamilton syringe in the automated injection apparatus.

Materials

Bone Wax	Surgical Specialities	903	Sterile and malleable beeswax and isopropyl palmitate
Cotton tip applicators	Puritan	25-806 10WC	Sterile long stemmed cotton tip applicators
Suture	Covidien	SP5686G	45 cm swedged 5-0 monofilament polypropylene suture
Skin Glue	Vetbond	1469SB	Skin safe wounds adhesive
Stereotaxic Unit
Mouse Gas Anesthesia Head Holder	Kopf	Model 923-B	Mouth bar with teeth hole and nosecone
Non-Rupture Ear Bars	Kopf	Model 922	Ear bars suitable for mouse applications
Stereotaxic Instrument	Kopf	Model 940	Base plate, frame and linear scale assembly with digital readout monitor
Injector
Injector Needle and syringe	Hamilton	80366	26 gauge needle, 51 mm needle length and 10 μL volume syringe
Legato 130A automated Syringe Pump	KD Scientific	P/N: 788130	Programmable touch screen base with automated injector
Anesthesia Machine
SomnoSuite Low-Flow Digital Vaporizer	Kent Scientific	SS-01	Digital anesthesia machine