Tumor growth

Examination of tumor growth was done as described previously¹⁴. Cells (1E6) suspended in HBSS were injected into the mammary fat pad of eight-week-old female NOD SCID mice. Six mice per group were used and the experiment was repeated one time with the independent passage of cells. Investigators were not blinded to any groups. Tumor growth was monitored by orthogonal measurements using Vernier caliper at a 3-day interval. Statistical significance was determined by two-way ANOVA followed by Dunnett’s post hoc test for tumor growth at respective time points and error bars represent SEM. Tumors were removed by survival surgery when control tumors reached a mean tumor diameter of 10 mm. Resected tumors were formalin-fixed paraffin-embedded. Eight weeks after tumor removal, the mice were euthanized, and the lungs were evaluated for metastasis. Lung metastases were enumerated using a Nikon StereoZoom microscope. The animal studies have been conducted in accordance with the approval and guidelines of the Institutional Animal Care and Use Committee (IACUC) of The University of Alabama at Birmingham.