Osteosarcoma xenograft models

Animal experiments were approved by the Institutional Animal Care Utilization Committee (IACUC) of Albert Einstein College of Medicine. 143B/shSkp2 or 143/shctrl cells were grown to 80% confluence and implanted orthotopically into the proximal tibia of SCID mice at 5 × 10⁵ cells (N = 8 animals in each group). Amputation of tumor-bearing legs of all mice was performed when any tumor reached 1.5 cm in diameter. Tumor tissues were harvested, measured, weighed and fixed in 10% formalin. Mice were then raised for two additional weeks after amputation and euthanized according to an IACUC-approved protocol. The lungs were harvested, weighed, and fixed in Bouin’s solution (Sigma-Aldrich Co., St. Louis, MO) to visualize and count surface nodules.

1 × 10⁶ 143B cells were injected into SCID mice through the tail vein. Animals were randomized into 2 groups (n = 8 per group): 0.1% DMSO or FKA (200 mg/kg per day). Treatment was initiated three days after injection of tumor cells. After 28 days, the animals were sacrificed according to an IACUC-approved protocol. The lungs were harvested, weighed, and fixed in Bouin’s solution.