How to Make a Liver Metastasis Model by Intrasplenic Inoculation of Tumor Cells in mouse?
(Protocol for a liver metastatic 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. 30-gauge needles
6. A 1-cc syringe
7. Tumor cells
Anesthetics:
Methoxyflurane
Methods:
The spleen is located in the left dorsocranial part of the abdominal cavity, its dark red color and elongated form makes it easy to be seen through the skin in nude mice. Inoculation of tumor cells into the spleen has proven to be a useful route to produce liver metastases.
1. Anesthetize a nude mouse with methoxyflurane inhalation.
2. Place the mouse in the right lateral decubitus position.
3. Scrub the left flank with Betadine followed by 70% alcohol.
4. Incise the skin and abdominal wall longitudinally (parallel to the spine) in the left flank for 0.5-1 cm.
5. Exteriorize the spleen and stabilize it gently.
6. Insert a 30-gauge needle on a 1-cc syringe into the parenchyma of the spleen for 3-4mm.
7. Inject 50 µl the material slowly. A visible pale wheal indicates a successful injection.
8. Retract the needle.
9. Place a Q-tip over the injection site for 30 seconds to prevent bleeding and spillage of material.
10. Return the spleen to the peritoneum.
11. Close the wound with wound clip in one layer.
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