Abstract: Objective To explore an efficient and reliable method for establishing an animal model of bone metastasis of tumors. Methods Male C57BL/6J mice were randomized into 4 equal groups to receive injections of normal saline or prostate cancer RM-1 cell suspension via the femoral artery or the external iliac artery, and breast cancer 4T1-luc cells were injected in 15 female BALB/c mice via the femoral artery. The operation time, postoperative survival rate, and the tumor formation rates in the mice with two different injection methods were compared. The tumor metastasis in the mice was evaluated with in vivo imaging. Results The mean time for hemostasis time was 2.53±1.75 min in mice receiving tumor cell injection via the femoral artery, significantly shorter than that in mice with injections via the external iliac artery (4.70 ± 1.63 min; P<0.05); the mean operation time was 14.67±2.16 min and 22.47±3.50 min in the two groups, respectively (P<0.05). At 21 days after the operation, the survival rate was 93.3% in femoral artery injection group, significantly higher than that in external iliac artery injection group (66.7%; P<0.05). The tumor metastasis rate was 100% in both groups. Conclusion Injection of the tumor cells via the femoral artery is more suitable for establishing mouse models of bone metastasis of cancers.