How to make a bone metastasis model by intracardiac injection in mouse?
(Protocol for a bone metastasis cancer model in mouse)
Materials:
1. 70% ethanol wipes and Betadine swabs
2. Q-tips
3. 30-gauge needles
4. A 1-cc syringe
5. Tumor cells
Anesthetics:
Pentobarbital sodium
Methods:
Inoculation of tumor cells into the left heart allows them to be distributed to all the organs of the body with the blood flow. This is particularly useful for studying the specific organotropism of tumor cells. Some selective cell lines, such as selected A375 melanoma cell line, tends to metastasize to bone after injected into left heart.
1. Anesthetize a mouse by intraperitoneal injection with 60-90mg/kg body weight of pentobarbital sodium.
2. Scrub the anterior chest wall with Betadine followed by 70% alcohol.
3. Insert a 30-gauge needle attached to a 1-cc syringe into the second intercostal space 3 mm to the left of the sternum and aimed centrally. Pull back slightly on the plunger when slowly advancing the needle, continuous entrance of red oxygenated blood into the transparent needle hub indicates proper positioning of the needle into the left heart.
4. When blood appears in the syringe, stop advancing the needle and fix the position of the needle steady.
5. Inject the materials from the syringe into the left heart slowly over a 20-40 second period.
6. Retract the needle, position the mouse laterally and keep the mouse warm to recover.
If you want to see the live demos of this specific technique by mouse work experts, please visit Video Training Courses.