Subcutaneous Xenograft Mouse Model

HMVP2 cells and spheroids were cultured for three days. Following trypsinization, cells were washed twice, re-suspended in a concentration of 2 × 10⁶ cells mixed with 200 μl 45% RPMI medium + 5% FBS + 50% matrigel membrane matrix (BD) and then the mixture was injected subcutaneously (SC) in a dorso-caudal location of two month old male FVB/N mice (N = 10). Mice were slightly sedated with isofluorane prior to injection. Similarly, approximately 200 spheroids resulting from an initial culture of 2 × 10⁵ cells were washed and re-suspended in fresh 45% RPMI medium + 5% FBS + 50% matrigel (200 μl total volume) and then injected subcutaneously (SC) in a second set of mice (N = 10). Mice were observed daily for tumor formation and small SC masses (~5 mm in diameter) were seen after two weeks of injection. Tumor masses were then measured twice a week with a digital caliper and tumor volume was calculated using the formula, tumor volume = ½ length × width² [60 (link)].