Creation of a murine orthotopic hepatoma model with intra-abdominal metastasis

To create an orthotopic hepatoma model with local metastasis monitored with ultrasound could be created as a platform for testing new treatments.

Hepatoma accounts for 25% of liver tumors in children with poor overall survival. Intraabdominal metastasis are present in 35% of patients at time of diagnosis. We hypothesized that an orthotopic tumor model with local metastasis could be created as a platform for testing treatment modalities and could be monitored with ultrasound.

One million human hepatoma cells (Hep3B) were injected into the left lobe of the liver of immunocompromised mice. Tumor volume was monitored with high frequency-ultrasound until it reached 1,000mm(3). At that time animals were sacrificed and examined for gross metastatic disease. Tumor sections were analyzed with hematoxylin and eosin (H&E) staining.

Tumor formed in 8/15 mice. The tumor was detected as small as 19.59mm(3) on ultrasound. Of the forming tumors, tumor size was 145±177.93mm(3) at 60 days post-injection, 665±650.39mm(3) at 67 days, and reached >1000mm(3) by 76.6±9.9 days. At necropsy, four mice (50%) had tumor only within the liver, four (50%) had additional tumors in omentum, pelvis and peritoneum. H&E showed tumor within the normal liver parenchyma, with multiple mitotic figures, small areas of necrosis, and hemorrhage within the tumor.

We have successfully established an orthotopic hepatoma murine model, with a local metastatic rate of 50%. Non-invasive tumor monitoring is feasible via ultrasound.