Nude Mice Xenograft Tumor Model

Eight-week-old female nude (nu/nu) mice (UCSD breeding colony) were housed in pathogen-free conditions. Tumor cells were washed in PBS, injected (2 × 10⁶ cells in 100 μl of PBS) subcutaneously into right and left flanks of nude mice, and tumor volume (length × width²/2) determined by Vernier caliper measurements over 24 d. Orthotopic tumor growth was initiated by surgical implantation (0.4 × 10⁶ cells in 7 μl of growth factor-depleted Matrigel) within the bursal region surrounding one ovary as described (12 (link)). Primary tumor weight was determined following euthanasia upon dissection. Fluorescent images of the intra-abdominal cavity and internal organs were acquired using an OV100 Small Animal Imaging System (Olympus). Blood was collected by heart puncture following euthanasia, samples were centrifuged, and serum was stored at −80°C. The UCSD Institutional Animal Care and Use Committee approved all mouse procedures.