Turrini P, Sasso R, Germoni S, Marcucci I, Celluci A, Di Marco A, Marra E, Paonessa G, Eutropi A, Laufer R, Migliaccio G, Padron J
Department of Pharmacology, IRBM P. Angeletti Merck Research Laboratories, Pomezia, Italy.
Transplant Proc. 2006 May;38(4):1181-4. doi: 10.1016/j.transproceed.2006.02.149.
The development of a small animal model for hepatitis C virus (HCV) infection is a critical issue for the development of novel anti-HCV drugs. To this aim, we have tried many different approaches for generating mice carrying humanized liver. Main efforts were focused on the transplantation of human hepatocytes into immunocompromised mice (SCID-/-, Bg-/-) carrying a genetic lethal liver disease (Alb-uPA). Survival of homozygotic animals should largely depend on early transplantation with healthy hepatocytes. In parallel to establishing a colony of Alb-uPA/SCID/Bg mice, we developed a microsurgical procedure for intrasplenic xenotransplantation of healthy hepatocytes in 1-week-old mice. So far, we generated several chimeras by xenotransplanting human hepatocytes in Alb-uPA+/+/SCID-/-/Bg-/- mice at 1 week after birth. In a first step, identification of successfully engrafted animals is possible by quantification of human serum albumin and human alpha 1 antitrypsin in mouse sera. Additional preliminary histomorphological analysis of liver sections from chimeric animals was also carried out. One of the mice was transiently infected with HCV, reaching viremia levels of approximately 10(5) genomes/mL. However, the efficiency of this system to generate chimeric mice is still very limited. We are currently exploring the use of more robust models of hepatic disease. Moreover, we have been also exploring novel strategies for the generation of chimeric mice by xenotransplanting human adult stem cells, instead of human hepatocytes, at preimmune stages of development.
开发丙型肝炎病毒（HCV）感染的小动物模型是新型抗HCV药物研发的关键问题。为此，我们尝试了多种不同方法来培育携带人源化肝脏的小鼠。主要工作集中于将人肝细胞移植到携带遗传性致死性肝病（Alb-uPA）的免疫缺陷小鼠（SCID-/-、Bg-/-）体内。纯合动物的存活在很大程度上应依赖于早期移植健康肝细胞。在建立Alb-uPA/SCID/Bg小鼠群体的同时，我们开发了一种显微外科手术，用于在1周龄小鼠的脾脏内进行健康肝细胞的异种移植。到目前为止，我们通过在出生后1周将人肝细胞异种移植到Alb-uPA+/+/SCID-/-/Bg-/-小鼠体内，培育出了几只嵌合体小鼠。第一步，可以通过定量小鼠血清中的人血清白蛋白和人α1抗胰蛋白酶来鉴定成功植入的动物。还对嵌合体动物肝脏切片进行了额外的初步组织形态学分析。其中一只小鼠被HCV短暂感染，病毒血症水平达到约10(5) 基因组/mL。然而，该系统生成嵌合体小鼠的效率仍然非常有限。我们目前正在探索使用更可靠的肝病模型。此外，我们也一直在探索在发育的免疫前阶段通过异种移植人成体干细胞而非人肝细胞来生成嵌合体小鼠的新策略。
