Oxygen-mediated enhancement of primary hepatocyte metabolism, functional polarization, gene expression, and drug clearance
Oxygen-mediated enhancement of primary hepatocyte metabolism, functional polarization, gene expression, and drug clearance
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DOI：
10.1073/pnas.0906820106
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发表时间：
2009-09-15
影响因子：
11.1
通讯作者：
Nahmias, Yaakov
中科院分区：
文献类型：
--
作者：
Kidambi, Srivatsan;Yarmush, Rubin S.;Nahmias, Yaakov
肝脏是异生物质化合物代谢的主要场所，这是由于其丰富的I/II相代谢酶水平。随着药物开发成本上升至超过4亿美元/药物，迫切需要开发严格的肝脏代谢模型，用于药物清除和肝毒性的临床前筛选。在这里，我们提出了一个微环境中，原代人类和大鼠肝细胞保持高水平的代谢能力，没有一个长的适应期。我们证明，肝细胞和内皮细胞在95%氧气下接种在无血清培养基中的共培养物保持功能性顶端和基底极性，高水平的细胞色素P450活性，以及与新鲜分离的肝细胞相当的基因表达谱。这些含氧共培养物显示出预测快速和缓慢清除药物的体内药物清除率的显著能力，R-2为0.92。此外，由于含氧共培养物的代谢功能在一夜之间稳定，因此可以在其他培养方法之前几天甚至几周进行临床前测试，从而显著降低相关的劳动力和成本。这些结果很容易扩展到其他文化配置，包括三维文化，生物反应器的研究，以及微加工共培养。
The liver is a major site for the metabolism of xenobiotic compounds due to its abundant level of phase I/II metabolic enzymes. With the cost of drug development escalating to over $400 million/drug there is an urgent need for the development of rigorous models of hepatic metabolism for preclinical screening of drug clearance and hepatotoxicity. Here, we present a microenvironment in which primary human and rat hepatocytes maintain a high level of metabolic competence without a long adaptation period. We demonstrate that co-cultures of hepatocytes and endothelial cells in serum-free media seeded under 95% oxygen maintain functional apical and basal polarity, high levels of cytochrome P450 activity, and gene expression profiles on par with freshly isolated hepatocytes. These oxygenated co-cultures demonstrate a remarkable ability to predict in vivo drug clearance rates of both rapid and slow clearing drugs with an R-2 of 0.92. Moreover, as the metabolic function of oxygenated co-cultures stabilizes overnight, preclinical testing can be carried out days or even weeks before other culture methods, significantly reducing associated labor and cost. These results are readily extendable to other culture configurations including three-dimensional culture, bioreactor studies, as well as microfabricated co-cultures.