【Abstract】Objective To explore the advantages and disadvantages of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish an orthotopic implantation model, and to provide technical reference for the establishment of an orthotopic implantation model. Methods A total of healthy Kunming mice were divided into four groups. Direct injection H22 cells was used in group A. Direct injection H22 ascitic cells was used in group B. Transplantation of tissues was used in group C. Direct injection was saline used in group D. The activity and weight change of each group were observed regularly. The survival time of the four groups of mice was recorded. Liver tumor formation, tumor size, abdominal organ adhesion degree and metastasis of the mice were observed. B-ultrasound imaging was performed to detect the concentration of alpha fetoprotein (AFP), abnormal prothrombin (DCP) and HE staining to observe liver histopathological changes. Results The mice in groups A, B and C were (3.36±0.44) min, (3.30±0.41) min and (5.68±0.65) min, respectively. After 25 d of modeling, the rate of mice in groups A, B and C was 100.0%, the severe abdominal adhesion in group A was 40.0%, 60.0% in group B, and neither group C or D. The rates of ascites in groups A, B and C were 40.0%, 100.0% and 0.0%, respectively; the rates of abdominal wall tumors were 30.0%, 60.0% and 0.0%, respectively. Mice in group B also had liver metastasis 40.0%). B ultrasound imaging, detection of serum AFP and DCP levels, histopathological results showed that mice in group A, B, C had smooth liver margins, uneven echo and slightly lower echo mass, maintaining AFP and DCP high secretion; large number of inflammatory cells and tumor cells. Conclusion H22 cells and H22 ascitic cells have the advantages of simple operation and high survival rate. transplantation of tissues has the advantages of less metastasis and light abdominal adhesion. All three methods can establish an orthotopic transplantation model of HCC, which can be an ideal model to evaluate the effect of drugs against HCC.