Modeling Orthotopic Bladder Tumor in Mouse Model: A Procedure for Intravesical Administration of Cancer Cells into Murine Bladder

Once a female mouse has been anesthetized, provide hydration by an intraperitoneal injection of Hartmann's solution at 0.1 milliliters per 10 grams of body weight. Repeat this every one or two hours during the anesthesia.

Next, apply sterile ophthalmic ointment to both eyes. Reapply as needed. Then, position the mouse supine on paper towels over a heat pad to maintain body temperature. Next, secure the hind legs with tape. Now, apply gentle pressure to the lower abdominal region and collect urine into a 1.5-milliliter tube.

Next, load a 1-milliliter syringe with sterile PLL and attach a 24-gauge IV catheter with the needle stylet removed. Apply lubricant to the tip of the catheter. Then, insert the catheter into the urethra and, using forceps, guide it to the bladder. Stop when resistance is felt. Then, slowly eject 50 microliters of PLL at a rate of 10 microliters every 20 seconds to avoid vesicoureteral reflux.

Now, leave the catheter in the bladder for 20 minutes with a stopper to prevent outflow. After 20 minutes, remove the catheter and vacate the bladder of any contents by gently pressing on the lower abdomen. Then, using a 1-milliliter syringe, flush any remaining contents out of the catheter.

Now, mix the MB49-PSA cells thoroughly by pipetting and load them into a 1-milliliter syringe. Attach the catheter and lubricate it as before. Insert the catheter into the bladder as before and commence with injecting the cells using a gradual ejection of 10 microliters per 20 seconds. Then, wait one hour.

After an hour, remove the catheter and vacate the bladder of any contents. Then, release the mouse. Position it on its ventral side and revive it with an injection of atipamezole. Now, monitor the mouse until it regains sternal recumbency and then return it to its home cage.