建立人鼠嵌合肝动物模型的实验研究  

Establishment of an animal model of tolerant rats with chimeric human liver

作　　者：蒋黎[1] 毛青[1] 王宇明[1] 李俊刚[1] 刘俊[1] 

机构地区：[1]第三军医大学西南医院全军感染病研究所,重庆400038

出　　处：《第三军医大学学报》2005年第9期856-859,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 2 71173)~~

摘　　要：目的　人胎肝细胞移植到具有正常免疫活性的大鼠体内是否不需要免疫抑制剂而能够存活。方法　Wistar大鼠出生前宫内腹腔注射人胎肝细胞,诱导胎鼠对人胎肝细胞产生免疫耐受,出生后2 4h内经脾移植人胎肝细胞。结果采用免疫印迹法(Westernblotting)、逆转录PCR(reversetranscriptionPCR ,RT PCR)及免疫组织化学等方法,于不同时相点检测实验大鼠血清人白蛋白、肝组织中人白蛋白mRNA及人肝细胞型细胞角蛋白CK18。结果于4、6、8周龄大鼠血清中测出人白蛋白;4、6、8、10周龄大鼠肝组织中测出人白蛋白mRNA ;4、6、8、10、12周龄大鼠肝组织中有CK18的表达。结论　采用诱导胎鼠出生前对人胎肝细胞产生免疫耐受,出生后移植人胎肝细胞的方法,使移植到具有正常免疫活性大鼠体内的人胎肝细胞,不需免疫抑制剂而能够存活,并且保持人肝细胞的特性,产生人白蛋白,维持近12周。初步建立人鼠嵌合肝动物模型。Objective To determine whether genetically normal immunocompetent rodent hosts could be manipulated to accept human fetal hepatocyte transplants and survive without immunosuppression. Methods Tolerance to human hepatocytes was established by injection of human fetal hepatocytes into the peritoneal cavities of fetal Wistar rats. Corresponding cells were subsequently transplanted into newborn rats via intrasplenic injection within 24 h after birth to establish tolerant rats with chimeric human liver. Results Transplanted human fetal hepatocytes in rat livers were visualized at different time with different assays after birth. Human albumin protein was detected in rat serum by Western blotting from week 4 to 8. Human albumin mRNA was detected in rat livers by RT-PCR from week 4 to 10, and cell keratin from human hepatocyte of cytokeratin 18 was also detected in rat livers by immunohistochemical staining from week 4 to 12. Conclusion Human fetal hepatocytes could be introduced into an immunocompetent rats at the appropriate time during fetal development, which could be rendered tolerant and serve as suitable hosts to human hepatocyte transplants after birth without either genetic or pharmacological generalized immunosuppression, and the survival time of functional human hepatocytes was 12 weeks.

关 键 词：嵌合 肝细胞移植 动物模型/鼠 

分 类 号：R-332[医药卫生] R617