The Primary Establishment Of Cryopreserved Patient-derived Hepatocellular Carcinoma Tissue Xenograft Subcutaneously

Object:The primary establishment and application of cryopreserved patient-derived hepatocellular carcinoma tissue xenograft subcutaneous.Materials and method:Divid the 5 cases of primary hepatocellular carcinoma which are obtained from liver surgery into two prats. Part of the cases of liver cancer can be divided into A, B, C, D four groups. After preserved 7days、30 day、90 day and 180 days successively,these tissues are used for the establishment subcutaneous transplantation of nude mice. Another part which is soaked in formalin is used for paraffin embedding, HE-staining and pathological observation. Record the formation rate, growth rate and the final tumor volume of the subcutaneous tumor in the nude mice. Through the comparisons of macroscopic character and pathomorphologic of tumor, and the expression of immunohistochemical Hep Par 1, AFP and CD34 in each group can we make a preliminary evaluation of this model.Result:Postoperative pathological diagnosis of 5 cases liver cancer were primary hepatocellular carcinoma. The freez-thawed tissues of the A groups,B groups,C groups,D groups were transplanted into the 60 nude mice subcutaneously totally, and there were 51 tumor-burdened nude mice. The tumor formation rate was 85%. The group D has longer incubation than group A, but the subcutaneous tumor growth of each group has no significant difference (P=0.954>0.05). The final tumor volume of each group is 867.24±29.81,889.30±13.88,862.10±29.41, 856.30±25.25 (mm3) and the result has no statistic significance. The patient-derived hepatocellular carcinoma tissue could growth stably after cryopreservation and anabiosisi. The transplanted subcutaneous tumor cell had irregular arrangement and the cell nucleus show different heteromorphism. The immunohistochemical results of Hep Parl, AFP and CD34 were all positive. The morphology and the biological characteristics remained as same as the original generation.Conclusion:The subcutaneous xenograft model of cryopreserved patient-derived hepatocellular carcinoma tissue is successively established. The freez-thawed tissue of patient-derived hepatocellular carcinoma could use for building the xenogrft model. Although the experiment found that the nude mice in group D has longer incubation than group A, but the formation rate and the growth rate has no significant difference and the transplanted tumor can keep the biological characteristics of the original generation. All the result show that the establishment of cryopreserved patient-derived hepatocellular carcinoma tissue xenograft subcutaneously could be successful, and that will make a contribute to the preservation of liver cancer and the establishment of hepatocellular carcinoma database.