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

1. Prepare Mice for Tumor Cell Injection

<ol>
	<li>Anesthetize a mouse by intraperitoneal (i.p.) injection of ketamine/xylazine cocktail (ketamine 100 mg/kg, xylazine 10 mg/kg).</li>
	<li>Confirm complete anesthesia by pinching the animal&#39;s feet and observe lack of response.</li>
	<li>Place the mouse on a glass p...

brain metastasis mouse model: a method to generate brain metastasis mouse model by intracarotid cancer cell injection

Source: Zhang, C. et al. <a href="https://www.jove.com/video/55085" target="_blank">Intracarotid Cancer Cell Injection to Produce Mouse Models of Brain Metastasis.</a> J. Vis. Exp. (2017)
This video demonstrates the establishment of a brain metastasis mouse model by intracarotid injection of fluorescently labeled cancer cells. The mouse model can be used to study the effects of novel therapeutics and the underlying physiological mechanisms.

Brain Metastasis Mouse Model: A Method to Generate Brain Metastasis Mouse Model by Intracarotid Cancer Cell Injection

Source: Zhang, C. et al. Intracarotid Cancer Cell Injection to Produce Mouse Models of Brain Metastasis. J. Vis. Exp. (2017)
This video demonstrates the ...

In brain metastasis, cancer cells from other affected organs, or the primary site of origin, spread into the central nervous system to form secondary tumors. To generate a mouse brain metastasis model, prep an anesthetized mouse in the supine position. Make a small incision in the neck region.
Dissect the muscles to expose one of the underlying carotid arteries. Carotid arteries are a pair of major blood vessels in the neck, supplying blood to the brain and head. Separate the carotid artery from the adjacent vagus nerve. Next, place suture knots around the proximal and distal ends of the artery.
Tighten the proximal knot to obstruct the blood flow into the target injection site. Then, place a buffer-soaked cotton ball under the target injection site to immobilize and stretch the artery for additional support. Take a syringe filled with green fluorescent protein or GFP-labeled cancer cell suspension.
Gently inject the cell suspension into the lumen of the carotid artery. Retract the needle and tighten the distal knot to prevent leakage of cancer cells. Lastly, suture the incision. Allow the mouse to recover. Over time, cancer cells begin to migrate and form secondary tumors in the brain, confirming the successful establishment of a brain metastasis model.

brain-metastasis-mouse-model-method-to-generate-brain-metastasis

Research

JoVE Encyclopedia of Experiments

Cancer Research

Cancers of the Nervous System

In vivo studies

Modeling Brain Metastases Through Intracranial Injection and Magnetic Resonance Imaging

Metastatic spread of cancer is an unfortunate consequence of disease progression, aggressive cancer subtypes, and/or late diagnosis. Brain metastases are particularly devastating, difficult to treat, and confer a poor prognosis. While the precise incidence of brain metastases in the United States remains hard to estimate, it is likely to increase as extracranial therapies continue to become more efficacious in treating cancer. Thus, it is necessary to identify and develop novel therapeutic approaches to treat metastasis at this site. To this end, intracranial injection of cancer cells has become a well-established method in which to model brain metastasis. Previously, the inability to directly measure tumor growth has been a technical hindrance to this model; however, increasing availability and quality of small animal imaging modalities, such as magnetic resonance imaging (MRI), are vastly improving the ability to monitor tumor growth over time and infer changes within the brain during the experimental period. Herein, intracranial injection of murine mammary tumor cells into immunocompetent mice followed by MRI is demonstrated. The presented injection approach utilizes isoflurane anesthesia and a stereotactic setup with a digitally controlled, automated drill and needle injection to enhance precision, and reduce technical error. MRI is measured over time using a 9.4 Tesla instrument in The Ohio State University James Comprehensive Cancer Center Small Animal Imaging Shared Resource. Tumor volume measurements are demonstrated at each time point through use of ImageJ. Overall, this intracranial injection approach allows for precise injection, day-to-day monitoring, and accurate tumor volume measurements, which combined greatly enhance the utility of this model system to test novel hypotheses on the drivers of brain metastases.

Intracranial brain metastasis modeling is complicated by an inability to monitor tumor size and response to treatment with precise and timely methods. The presented methodology couples intracranial tumor injection with magnetic resonance imaging analysis, which when combined, cultivates precise and consistent injections, enhanced animal monitoring, and accurate tumor volume measurements.

Metastatic spread of cancer is an unfortunate consequence of disease progression, aggressive cancer subtypes, and/or late diagnosis. Brain metastases are ...

JoVE Journal

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous substances into the brain. These exogenous substances may induce injurious impact or constitute therapeutic strategy. The most common delivery methods of exogenous substances into the brain are based on systemic deliveries, such as subcutaneous or intravenous routes. Although commonly used, these approaches have several limitations, including low delivery efficacy into the brain. In contrast, surgical methods that locally deliver substances into the CNS are more specific and prevent the uptake of the exogenous substances by other organs. Several surgical methods for CNS delivery are available; however, they tend to be very traumatic. Here, we describe a mouse infusion microsurgery technique, which effectively delivers substances into the brain via the internal carotid artery, with minimal trauma and no interference with normal CNS functionality.

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

The present protocol describes a mouse microsurgery infusion technique, which effectively delivers substances directly into the brain via the internal carotid artery.

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous ...

Neuroscience

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to intracranial delivery, intra-arterial administration of NSCs is less invasive and produces a more diffuse distribution of NSCs within the brain parenchyma. Further, intra-arterial delivery allows the first-pass effect in the brain circulation, lessening the potential for trapping of cells in peripheral organs, such as liver and spleen, a complication associated with peripheral injections. Here, we detail the methodology, in both mice and rats, for delivery of NSCs through the common carotid artery (mouse) or external carotid artery (rat) to the ipsilateral hemisphere after an ischemic stroke. Using GFP-labeled NSCs, we illustrate the widespread distribution achieved throughout the rodent ipsilateral hemisphere at 1 d, 1 week and 4 weeks after postischemic delivery, with a higher density in or near the ischemic injury site. In addition to long-term survival, we show evidence of differentiation of GFP-labeled cells at 4 weeks. The intra-arterial delivery approach described here for NSCs can also be used for administration of therapeutic compounds, and thus has broad applicability to varied CNS injury and disease models across multiple species.

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

Brain Metastasis Mouse Model: A Method to Generate Brain Metastasis Mouse Model by Intracarotid Cancer Cell Injection

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