This article describes the generation of a patient-derived urothelial cell carcinoma (UCC) mouse model to study cancer progression and metastasis. The procedure involves inoculating immunocompromised mice with human UCC cells through a series of precise steps to ensure successful engraftment.
For a patient-derived urothelial cell carcinoma or UCC mouse model generation, an immunocompromised mouse is inoculated with human UCC cells to study cancer progression and metastasis.
To begin xenografting, take an immunocompromised anesthetized female mouse. Shave the lower back of the mouse to expose its pelvic region. Now, place the mouse in the supine position with its exposed skin in contact with a dispersive pad that serves as a grounding electrode to complete the circuit during electrical stimulation.
Next, take an angiocatheter, a thin hollow tube, attached to a guidewire. Lubricate the surface of the catheter and insert it into the mouse's urethra to access its bladder. Place an active electrode on the guidewire to stimulate the bladder wall. This procedure helps rupture the mucopolysaccharide layer, a protective barrier of the bladder mucosa, and aids in the subsequent engraftment of tumor cells.
Now, gently remove the angiocatheter-guidewire assembly from the urethra. Take a second angiocatheter attached to a syringe containing the UCC cell suspension and insert it into the mouse's urethra. Inject the UCC cells into the mouse's bladder. Cancer cells interact with the bladder mucosa to form a tumor that may eventually spread to other tissues and organs.
To prepare the animal for UCC cell injection, use hair removal cream to shave the lower back of a 6- to 8-week old female NOD/SCID mouse. To administer UCC cells into the bladder, first, set up a monopolar electrocautery machine to a power of 4 watts. After anesthetizing the mouse, place it in supine position with its snout in an isoflurane nose cone and bare back firmly grounded on a dispersive electrode.
Lubricate a 24-gauge angiocatheter with lubricating jelly and then insert the angiocatheter gently but firmly through the mouse urethra. If catheter bends on entry, insert guide wire halfway into catheter to provide stability. Then, fully insert the 0.64-millimeter fixed core straight guidewire 1-millimeter past the end of the angiocatheter.
Hold the monopolar pin to the guidewire for 1 second, allowing for electrical irritation of the bladder mucosa. Attach a fresh sterile angiocatheter to 1-milliliter Luer-Lok syringe and draw up to 200 microliters of UCC cells. Remove guide wire and angiocatheter from the urethra and then insert angiocatheter with syringe of cells attached to it.
Inject 50 microliters of the cells to the mouse bladder, and to allow the cells to adhere to the bladder wall, wait a few seconds before removing the angiocatheter. Finally, remove the mouse from isoflurane nose cone and grounding pad and observe it for 1 hour following procedure for signs of distress.
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