Objective Total thyroidectomy of C57BL/ 6 and KM mice was performed by two different surgical Methods to verify the success of mouse hypothyroidism model modeling, and compared the success rate of different surgical Methods. Methods C57BL/ 6 and KM mice underwent total thyroidectomy by ligation ( operation method Ⅰ) or hemostasis (operation method Ⅱ), and the detailed operation processes were recorded. Serum TT3, TT4 and TSH levels detected by enzyme-linked immunosorbent assay, body weight, and hematoxylin-eosin ( HE)-stained neck tissue were compared before and after surgery to verify the model. Results Serum TT3 and TT4 levels were decreased (P< 0. 05) and TSH was increased (P< 0. 001) in both model groups. The 28-day postoperative survival rates were 40% and 60% in groups Ⅰ and Ⅱ, respectively, and 50% and 40% in KM mice. Body weights were significantly higher in both model groups compared with the sham control group. HE staining and microscopic observation showed that the cervical tissue in both strains was thyroid tissue, and the back membrane of the thyroid remained intact after isolation. Conclusions Both surgical method can induce hypothyroidism in C57BL/ 6 and KM mice; however, it is necessary to consider the anatomical relationship of the thyroid gland to the surrounding tissue, improve the proficiency of the surgical operation, prevent the occurrence of postoperative hypocalcemia and infection, and thus improve the survival rate of the model mice.