Waring Jeffrey F, Ciurlionis Rita, Jolly Robert A, Heindel Matthew, Gagne Gerry, Fagerland Jane A, Ulrich Roger G
Department of Cellular and Molecular Toxicology, Abbott Laboratories, Abbott Park, IL 60064-6123, USA.
Toxicol In Vitro. 2003 Oct-Dec;17(5-6):693-701. doi: 10.1016/s0887-2333(03)00102-4.
In vitro human hepatocyte cultures are a key tool in the investigation of xenobiotic toxicity and metabolism. In most in vitro hepatocyte studies, the cells are allowed to adhere to an extracellular matrix, such as collagen. Unfortunately, the ability of freshly isolated hepatocytes to adhere to collagen varies from donor to donor. We used microarray analysis to determine what gene expression differences exist between hepatocytes in suspension and hepatocytes attached to collagen. Results from different donors showed a considerable difference in gene expression patterns between the two hepatocyte populations. In addition, we also compared the gene expression profiles of hepatocytes in culture with liver tissue. The results showed that both hepatocytes in suspension and hepatocytes attached to collagen display significant gene expression differences compared with liver tissue. Finally, we show that both populations of hepatocytes are responsive to dexamethasone and regulate some of the same genes. Overall, our results suggest that either significant gene expression changes occur in isolated hepatocytes or that suspended and attached cells represent different populations of hepatocytes found in intact livers.
体外人肝细胞培养是研究外源化合物毒性和代谢的关键工具。在大多数体外肝细胞研究中，细胞被允许附着于细胞外基质，如胶原蛋白。不幸的是，新鲜分离的肝细胞附着于胶原蛋白的能力因供体而异。我们使用微阵列分析来确定悬浮肝细胞和附着于胶原蛋白的肝细胞之间存在哪些基因表达差异。来自不同供体的结果显示，这两种肝细胞群体之间的基因表达模式存在相当大的差异。此外，我们还比较了培养的肝细胞与肝组织的基因表达谱。结果表明，与肝组织相比，悬浮肝细胞和附着于胶原蛋白的肝细胞均表现出显著的基因表达差异。最后，我们表明这两种肝细胞群体均对地塞米松有反应，并调节一些相同的基因。总体而言，我们的结果表明，要么在分离的肝细胞中发生了显著的基因表达变化，要么悬浮细胞和附着细胞代表了完整肝脏中发现的不同肝细胞群体。
