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Cell Biology JoVE (Open Access) Citable · DOI

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

DOI: 10.3791/51123-v
What you'll learn
  • ✓ Perform percutaneous ultrasound-guided bladder xenograft inoculation in mice
  • ✓ Separate bladder wall layers and deliver cancer cells intramurally
  • ✓ Establish orthotopic bladder tumors with minimal surgical morbidity
  • ✓ Troubleshoot injection placement and cell viability in xenograft models
Protocol

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

Difficulty
advanced
Total time
~45-60 min per mouse (including ultrasound setup and injection)
Model organism
Mouse (strain not specified; orthotopic xenograft model)
Biosafety
BSL-2

Steps

1
Prepare cell lines for bladder xenograft inoculation

Culture and prepare primary invasive bladder cancer cells at appropriate concentration and viability for percutaneous injection. Ensure cells are in suspension and ready for loading into injection apparatus.

▶ 01:37
2
Prepare animals and ultrasound imaging setup

Anesthetize mice, position for supine imaging, and establish ultrasound guidance system. Prepare sterile injection field and verify bladder visualization on ultrasound.

▶ 02:54
3
Separate bladder wall layers and deliver cancer cells

Use ultrasound guidance to percutaneously access the bladder, identify distinct wall layers, and inject cancer cells into the intramural compartment with high precision. Monitor needle placement throughout injection.

▶ 04:14
4
Evaluate injection success and tumor establishment

Assess representative results from successful injections, including verification of intramural cell placement, absence of leakage, and confirmation of viable xenograft initiation on post-operative imaging.

▶ 05:54
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