Surgical Cisterna Magna Injection: A Method for Administering Tumor Cells Directly into Central Nervous System of Murine Model

Abstract

Source: Law, V. et al. A Murine Ommaya Xenograft Model to Study Direct-Targeted Therapy of Leptomeningeal Disease. J. Vis. Exp. (2021)

In this video, we describe the procedure to deliver human-derived tumor cells into the cisterna magna of a mouse’s brain. The orthotopic mouse model can be used to study metastatic spread of cancer to the cranial meninges.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Preparation of CTCs

1. Calculate the number of CTCs needed for injection and prepare a single-cell suspension at 1.0 × 10⁴ cells/μL in sterile phosphate-buffered saline (PBS). Place the cell suspension on ice or at 4 °C throughout the procedure.
   NOTE: When using cell lines that require trypsinization, be sure to wash cells twice with sterile PBS to remove trypsin. Perform cell count using a hemocytometer or an automated ....

Results

Figure 1: Injection of circulating tumor cells into the cisterna magna in a murine xenograft model to study leptomeningeal disease and central nervous system metastases. (A) An illustration showing the location of the cisterna magna and the CSF accessing site, in which CTCs are injected using .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Bead sterilizer 	Braintree Scientific Inc. (or any vendor) 	#GER 5287-120V 	Germinator 500
Buprenorphine Sustained-Release (Bup-SR) 	Zoopharm 		DEA controlled
Hamilton microliter syringes&nbsp;	Hamilton&nbsp;		10, 25, 50, and 100 &mu;L;<br/> 30G for cisterna magna injection
Phosphate-buffered saline (PBS)&nbsp;	Any vendor		0.1 mm Sterile-Filtered
Rodent Surgical Instruments (Scissors, Forceps)	Roboz Surgical Instrument (or any vendor)
Sterile blue paper/ drape covering&nbsp;	Any vendor
Sterile cotton sticks&nbsp;	Any vendor