Objective To explore the otherness of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish orthotopic implantation models of hepatocellular carcinoma in mice, and to provide a technical reference for the establishment of an orthotopic implantation model. Methods Healthy KM mice were divided into four groups: group A, direct injection of H22 cells; group B, direct injection of H22 ascitic cells; group C,transplantation of tissues; and group D, direct injection of saline. Activity and weight changes were observed regularly in each group and survival times were recorded. Liver tumor formation, tumor size, abdominal organ adhesion degree, and metastasis were observed in all groups. B-ultrasound imaging was performed, concentrations of alpha fetoprotein (AFP) and abnormal prothrombin (DCP) were detected, and liver histopathological changes were detected by hematoxylin and eosin staining. Results Mice molding operation time 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 modeling for 25 days, the rates of model formation in groups A, B, and C were all 100. 0%. Severe abdominal adhesions occurred in 40. 0% of mice in group A and 60. 0% in group B, but in no mice in group C or D. Ascites occurred in 40. 0%, 100. 0%, and 0. 0% and abdominal wall tumors in 30. 0%, 60. 0%,and 0. 0% of mice in groups A, B, and C, respectively, while 40. 0% of mice in group B also had liver metastasis. Bultrasound imaging, detection of serum AFP and DCP levels, and histopathological result showed smooth liver margins,uneven echo and slightly lower echo mass, maintained high AFP and DCP secretion, and large numbers of inflammatory cells and tumor cells in mice in groups A, B, and C. Conclusions At day 25, all three Methods can thus be used to establish orthotopic transplantation models of HCC. Among these, inj ection of cell suspensions demonstrated the advantage of simplicity in operation and the presence of multiple metastatic nodules within the liver, compared to transplantation of tumor tissue. Conversely, transplantation of tumor tissue showed the advantage of causing less impact on the abdomen and other organs when compared to inj ection of cell suspensions.