Yuan Jing, Liu Liegang, Shimada Masashi, Wang Aiguo, Ruhnke Maren, Heeckt Peter, Muller Andrea R, Nussler Natascha C, Neuhaus Peter, Nussler Andreas
Department General, Visceral and Transplantation Surgery, Charité Campus Virchow-Klinikum, Universitätsmedizin Berlin, Humboldt University of D-Berlin, Germany.
ALTEX. 2004;21 Suppl 3:3-11.
Within the past decade, tremendous progress has been made in the isolation and culture of human hepatocytes for drug metabolism and toxicology, which could potentially reduce the number of animal experiments performed. However, human hepatocyte cultures are still not widely used for preclinical drug testing, partly due to inconsistent supply and quality of human tissue. Thus, the aim of this study was to evaluate primary cultured human hepatocytes from different patients over a study period of 14 days, by assays that characterise cell quality and function. We found urea production and albumin synthesis in all cell cultures over at least 7 days. Cytochrome P4501A2, CYP2D6, and CYP3A4 protein expression was demonstrated by Western Blot analysis and CYP1A1/2 and CYP3A4 induction by 3-methylcholantrene, phenobarbital or rifampicin over 14 days. In addition, we saw that UDP-glucoronyltransferase activity was preserved in human hepatocytes over 2 weeks. In conclusion, we could show that primary human hepatocytes isolated from discarded liver tissue can consistently be kept in culture over a long time period and are therefore well suited for preclinical drug testing.
在过去十年中，人类肝细胞的分离和培养在药物代谢和毒理学方面取得了巨大进展，这有可能减少所进行的动物实验数量。然而，人肝细胞培养仍未广泛用于临床前药物测试，部分原因是人体组织的供应和质量不一致。因此，本研究的目的是通过表征细胞质量和功能的检测方法，在14天的研究期内评估来自不同患者的原代培养人肝细胞。我们发现所有细胞培养物中至少7天内都有尿素生成和白蛋白合成。通过蛋白质印迹分析证实了细胞色素P4501A2、CYP2D6和CYP3A4蛋白的表达，以及14天内3-甲基胆蒽、苯巴比妥或利福平对CYP1A1/2和CYP3A4的诱导作用。此外，我们还发现尿苷二磷酸葡萄糖醛酸基转移酶活性在人肝细胞中可保持2周。总之，我们可以证明，从废弃肝脏组织中分离的原代人肝细胞能够长期稳定地保持在培养状态，因此非常适合用于临床前药物测试。
