Murine CCA Liver Xenograft Model

All animal experiments were performed in accordance with a protocol approved by the Mayo Clinic Institutional Animal Care and Use Committee. Murine CCA cells (SB1-7) were harvested and washed in DMEM. Male C57BL/6 mice from Jackson Labs were anesthetized using 1.5–3% isoflurane. Under deep anesthesia, the abdominal cavity was opened by a 1 cm incision below the xiphoid process. A sterile cotton tipped applicator was used to expose the superolateral aspect of the medial lobe of the liver. Using a 27-gauge needle, 40 µL of standard media containing 1 × 10⁶ cells was injected into the lateral aspect of the medial lobe. Cotton tipped applicator was held over the injection site to prevent cell leakage and blood loss. Subsequently, the abdominal wall and skin were closed in separate layers with absorbable chromic 3–0 gut suture material.