Runge D, Runge D M, Jäger D, Lubecki K A, Beer Stolz D, Karathanasis S, Kietzmann T, Strom S C, Jungermann K, Fleig W E, Michalopoulos G K
Klinik für Innere Medizin I, Martin Luther Universität Halle-Wittenberg, Halle, 06097, Germany.
Biochem Biophys Res Commun. 2000 Mar 5;269(1):46-53. doi: 10.1006/bbrc.2000.2215.
Since human hepatocytes are available only in limited number, the development of a serum-free culture system for long-term cultivation of differentiated and functional hepatocytes is of great importance. Here we describe the culture of human hepatocytes in a chemically defined serum-free medium for up to 5 weeks. Cell morphology was assayed by light and electron microscopy and revealed a well-preserved cellular morphology. Marker proteins for epithelial and bile duct cells, cytokeratin (CK) 18 and 19, and liver-specific proteins, like phosphoenolpyruvate carboxykinase-2 (PCK2) and serum proteins, were expressed. Liver-enriched transcription factors CCAAT/enhancer binding protein alpha (C/EBPalpha) and hepatocyte nuclear factor-4 (HNF-4), cytokine and mitogen activated factors (nuclear factor kappa B) NFkappaB, and activator protein-1 (AP-1) were maintained and active for several weeks in our cultures. In summary, our serum-free culture system allows the culture of differentiated human hepatocytes for several weeks. It may serve as a model system for metabolic, pharmacologic-toxicologic studies, and studies on human pathogens under defined chemical conditions.
由于人肝细胞数量有限，因此开发一种用于长期培养分化且具有功能的肝细胞的无血清培养系统至关重要。在此，我们描述了在化学成分明确的无血清培养基中对人肝细胞进行长达5周的培养。通过光学显微镜和电子显微镜对细胞形态进行检测，结果显示细胞形态保存良好。上皮细胞和胆管细胞的标志物蛋白细胞角蛋白（CK）18和19，以及肝脏特异性蛋白，如磷酸烯醇丙酮酸羧激酶-2（PCK2）和血清蛋白均有表达。肝脏富集转录因子CCAAT/增强子结合蛋白α（C/EBPα）和肝细胞核因子-4（HNF-4）、细胞因子和丝裂原激活因子（核因子κB）NFκB以及激活蛋白-1（AP-1）在我们的培养物中持续保持活性数周。总之，我们的无血清培养系统能够对分化的人肝细胞进行数周培养。它可作为代谢、药物毒理学研究以及在特定化学条件下研究人类病原体的模型系统。
