Modeling Orthotopic Bladder Tumor in Mouse Model: A Procedure for Intravesical Administration of Cancer Cells into Murine Bladder
Transcripción
Intravesical instillation facilitates cancer cell delivery into the urinary bladder of a recipient animal. To begin, prep an anesthetized female mouse in the supine position on a heating pad to regulate its body temperature. Empty out the bladder of urine.
Next, assemble a catheter connected to a syringe containing poly-L-lysine or PLL solution. Lubricate the catheter tip to ensure smooth urethral insertion and gain access into the bladder. Following insertion, slowly inject the PLL solution into the bladder.
The slow delivery of PLL prevents vesicoureteral reflux – the backflow of urine from the bladder to the ureters and kidneys. Place a stopper on the catheter to prevent premature out-flow. The injected positively-charged PLL molecules interact and adhere to the negatively-charged glycosaminoglycan coating on the inner wall of the bladder.
Remove the catheter and vacate the bladder of unbound PLL solution and urine. Using another catheter-syringe assembly, slowly inject a chilled cancer cell suspension into the bladder to prevent vesicoureteral reflux and ensuing cell implantation in kidneys. Fix the stopper on the catheter.
The altered surface charge of the glycosaminoglycan layer enhances its electrostatic interaction with cancer cells, facilitating their adhesion. Remove the catheter and vacate the bladder of unadhered cancer cells and urine. Over time, the implanted cancer cells within the bladder proliferate to establish primary tumors.
