Olubuyide I O, Judah D J, Riley J, Neal G E
MRC Toxicology Unit, MRC Laboratories, Carshalton, Surrey, UK.
Br J Cancer. 1991 Mar;63(3):378-85. doi: 10.1038/bjc.1991.89.
The preparation of primary cultures of control and DHBV-infected duck hepatocytes from embryos and young ducklings is described. Cultures of both embryo and duckling hepatocytes secreted duck serum proteins. Cultures of hepatocytes established from ducklings maintained initial morphology for up to 3 weeks in culture and also exhibited high levels of metabolism of aflatoxin B1. Embryonic cell cultures rapidly lost ability to metabolise AFB1 and became overgrown by spindle-shaped cells. Both embryo and duckling cell cultures secreted infective DHBV, and had intracellular replicative forms of the virus. No integration of the virus into the duck genome was observed, and attempts to induce viral integration in the duckling hepatocytes using irradiation and aflatoxin B1 toxicity were unsuccessful. The results of the study lend further support to the suggestion that the rarity of liver cancer in DHBV-infected experimental ducks is related to an innate resistance of the hepatocytes to develop DHBV-DNA integration. Another possibility may be related to the lower oncogenic potential of the DHBV strain used for the study. However DHBV infected duckling hepatocytes would appear to offer a suitable material for studying viral replication and mechanisms of aflatoxin B1 toxicity during prolonged cell culture.
本文描述了从胚胎和幼鸭制备对照和DHBV感染的鸭肝细胞原代培养物的方法。胚胎和幼鸭肝细胞培养物均分泌鸭血清蛋白。从幼鸭建立的肝细胞培养物在培养中保持初始形态长达3周，并且还表现出高水平的黄曲霉毒素B1代谢。胚胎细胞培养物迅速丧失代谢AFB1的能力，并被纺锤形细胞过度生长。胚胎和幼鸭细胞培养物均分泌感染性DHBV，并具有病毒的细胞内复制形式。未观察到病毒整合到鸭基因组中，并且使用辐射和黄曲霉毒素B1毒性诱导幼鸭肝细胞中病毒整合的尝试未成功。该研究结果进一步支持了以下观点：DHBV感染的实验鸭中肝癌罕见与肝细胞对DHBV-DNA整合的先天抗性有关。另一种可能性可能与用于该研究的DHBV毒株的致癌潜力较低有关。然而，DHBV感染的幼鸭肝细胞似乎为研究长期细胞培养过程中的病毒复制和黄曲霉毒素B1毒性机制提供了合适的材料。
