How to Make an Orthotopic Model of Bladder Cancer in Mouse?
(Protocol for an orthotopic bladder 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 bladder lies ventrally to the colon in the dorsocaudal abdominal cavity, usually filled with urine, easy to be located.
1. Anesthetize a nude mouse with methoxyflurane inhalation.
2. Place the mouse on its back.
3. Scrub the lower abdomen wall with Betadine followed by 70% ethanol.
4. Incise the skin and peritoneum along the lower midline for about 1 cm.
5. Exteriorize the urinary bladder.
6. Pull it upwards gently with a smooth forceps to evacuate its urine content. Stabilize the bladder gently.
7. Insert a 30-gauge needle attached to a 1-cc syringe obliquely into the bladder wall. Inject 50 µL of tumor cell suspension slowly. A well-localized bleb indicates a successful injection.
8. Retract the needle.
9. Place a Q-tip over the injection site for about 30 second to prevent bleeding and spillage of material.
10. Return the bladder to the peritoneum.
11. Close the abdominal wall with wound autoclip in one layer.
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