4.10. Xenograft in Nude Mice

Tumorigenicity was assayed by the subcutaneous injection of 5 × 10⁶ cells of 786-O and 10⁶ of ACHN, A498 and CaKi-1 cell lines into the right flanks of 4-week-old female athymic nude mice. Tumorsphere tumorigenicity was measured by seeding 10⁴ cells as described in tumorspheres assay section, after 5 days tumorspheres were disaggregated with 0.025% trypsin-EDTA, resuspended in 50 µL of media and 50 µL of matrigel (Corning, Corning, NY, USA) and injected into the right flanks of 4-week-old female athymic nude mice. Animals were examined weekly. Tumor volume (mm³) was measured using calipers. All animal experiments were performed according to the experimental protocol approved by the IBIS and HUVR Institutional Animal Care and Use Committee (0309-N-15).