Diagnosis for hepatitis, development of a vaccine for hepatitis virus and development of a new drug for liver diseases using primary culture of human hepatocytes

原代培养的成年大鼠肝细胞维持许多肝脏功能，并以与体内肝细胞相同的方式对各种激素作出反应。此外，成熟的肝细胞在体外低细胞密度培养时，如EGF和肝细胞生长因子（HGF）等生长因子可以增殖。最近，我们发现低浓度的C <Cl_4>可引起培养的成年大鼠肝细胞的酶渗漏，这是由于细胞膜的通透性发生了变化，并证明了甘草酸苷和糖皮质激素对C <Cl_4>诱导的细胞酶渗漏有保护作用。因此，成年大鼠肝细胞的原代培养对肝脏疾病毒理学和新药开发具有重要意义。然而，尽管这些关于大鼠肝细胞的发现对理解人类肝脏学非常有用，但它们不能直接应用于人类肝脏。因此，有必要对原代培养的人肝细胞进行研究，并与大鼠肝细胞进行比较。特别是，由于人肝炎病毒具有严格的种特异性，因此利用人肝细胞来研究这种病毒的感染，特别是对非甲、非乙型肝炎病毒的鉴定至关重要。从肝活检标本中分离肝细胞，在无血清培养基中培养成单层。我们发现胰岛素、EGF和HGF均能诱导人肝细胞增殖，并且肝细胞的生长抑制非实质肝细胞的生长。人肝细胞原代培养对非甲、非乙型肝炎病毒的诊断、鉴定和疫苗的开发是有用的。

Adult rat hepatocytes in primary culture maintain many liver functions and respond to various hormones in the same ways as hepatocytes do in vivo. Moreover, mature hepatocyes can proliferate in vitro when cultured at low cell density with growth factors such as EGF and hepatocyte growth factor(HGF). Recently, we showsthat C <Cl_4> at low concentrations causes enzyme leakage from cultured adult rat hepatocytes due to a change in the permeability of the cell membrane and also demonstrate that glycyrrhyzin and glucocorticoid has a protective effect on enzyme leakage from the cell induced by C <Cl_4> . Thus, primary cultures of adult rat hepatocytes are a useful for toxicology and development of a new drug for liver disease. However, although these findings with rat hepatocytes must be very useful in understanding human hepatology, they cannot be applied to human liver directly. Therefore, it is necessary to study human hepatocytes in primary culture in comparison with those of rat liver. In particular, since human hepatitis virus has strict species specificity, it is essential to use human hepatocytes to study this virau infection, especially for the identification for non-A, non-B hepatitis virus. Hepatocytes were isolated from liver biopsy specimens and cultured in serum-free medium as monolayers. We found that proliferation of human hepatocytes can be induced by insulin EGF and HGF, and that growth of hepatocytes suppressed that of non-parenchymal liver cells in co-culture. Primary cultures of human hepatocytes are useful for diagnosis, identification and development of a vaccine for non-A, non-B hepatitis virus.