Bladder cancer (BC) is one of the most prevalent cancers of the genitourinary system. Animal models offer an important tool to research tumour initiation, development, and treatment processes. The researchers aim was to create an optimum orthotopic BC model which is predictable, reproducible, and convenient. The optimised orthotopic BC model was generated in male C57BL/6 mice utilising microsyringes to inoculate them with a murine BC cell line (MB449) (MB49). Anaesthetise mice were inoculated with an MB49 cell solution (10 µL) at about 5 × 106/mL. The full process of modelling was viewed and evaluated every 3 days for 21 days utilising HE staining and transabdominal ultrasonography (TUS). In this investigation, the model exhibited outstanding success rates for cancer formation (96.67%) and metastatic rate (89.66%). Compared to the control group (sham operation), mice in the modelling group showed serous cachexia, evident haematemesis and weight loss (all P<0.05). The lungs, liver, ureter and kidneys were discovered to have cancer metastases. Moreover, the average survival time (19.73±1.69 d) of modelling mice was substantially less than that of the control mice (P<0.05), which stayed alive. The researchers devised a method using microsyringes to inject murine BC cells into the bladder wall, generating a durable transplantable BC model in mice.