Establishment Of Animal Model In Nude Mice By Retransplantation Human Cholangiocarcinoma Tissue Into Subcutaneous And Observation

OBJECTION: To establish a xenografted model of hilar cholgangiocarcinom in nude mice METHOD: A fresh human hilar cholangiocarcinoma specimen was obtained from a patient under aseptic condition during operation, included primary and metastatic tissue. Using parts of primary tissue, divided it into inner ,middle, outer three parts, quickly cutting these into 1mm3 small pieces, with metastatic tissue, and randomly to establish primary(inner), primary(middle), primary(outer),and metastatic four groups animal models, and will retransplantate and observe from generation to next generation(total 3 generations) ;The tumor specimen was cut into pieces, cultured in 1640 medium, and subcutaneously transplanted into Balb/c- nu/nu mice. When the tumors reached 10 mm in diameter, or the host showed cachexia, the transplanted tumors were retransplanted into new mice. The mice were observed every 5 days, and the growth of tumor was recorded and growth curves were figured. The mice were sacrificed when they showed serious illness, and laparotomized to exminemetastatic tumors in liver and lungs. Direct dissemination method was used to affirm the karyotype; vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF2), proliferating cell nuclear antigen (PCNA) was examined with immunohistochemical methods. Flow cytometry was used to examine every period's percentage of cell cycle ; Carbohydrate antigen 242(CA242), carcinoembryonic antigen (CEA) were examined in radioimmunoassay.RESULT: The tumor was mobile, a litle stif and secreted amounts of mucus. HE staining showed a poorly differentiated adenocarcinoma structure, just like the human hilar cholangiocarcinoma specimen. The tumor was successfully passaged 3 times. The incubation period was shortened to 3-4 days and the tumor grew faster than the first passage. No liver or lung metastatic tumors were observed in all the mice VEGF, FGF2, PCNA were heavily expressed. Flow cytometry examination shows that the ratio of S phase fraction in metastatic tissue is higher than that in primary tissue. According to radioimmunoassay, the cellular concentrations of some tumor markers were as follow: the two markers were all negative in serum. Except the first passage growth curve, other passages were very similar. The third passage presented a steady growth status.CONCLUSION: The xenografted hilar chlgangiocarcinoma in nude mice model accords with human...