BEANS Laboratory, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan.
Lab Chip. 2012 Apr 24;12(10):1857-64. doi: 10.1039/c2lc40046d. Epub 2012 Mar 23.
Analysis of biliary metabolites is essential to predict pharmacokinetics and hepatotoxicity during drug development. In this paper, we present a hepatocyte culture configuration that enables the direct recovery of bile acid that accumulates in bile canaliculi by embedding the hepatocytes in a 3D micropatterned collagen gel substrate. We investigated the formation of bile canaliculi in hepatocytes embedded in circular microcavities of various sizes and made from collagen gel. Image analyses using fluorescently labeled bile acid revealed that the area of bile canaliculi in embedded hepatocytes in a microcavity of 60 or 80 μm in diameter was enlarged when compared with other sized microcavities and those of hepatocytes cultured using conventional hepatocyte sandwich cultures. We successfully recovered bile acid from the enlarged bile canaliculi of hepatocytes cultured in microcavities using a glass capillary and quantified the amount recovered. Using our approach, the direct recovery of biliary metabolites, using hepatocyte cultures with enhanced biliary excretion and geometrically enlarged bile canaliculi, may enable accurate screening of pharmacokinetics and drug-drug interactions against drug transporters.
对胆汁代谢物进行分析对于在药物开发过程中预测药代动力学和肝毒性至关重要。在本文中，我们提出了一种肝细胞培养结构，通过将肝细胞嵌入 3D 微图案胶原凝胶基质中，可以直接回收在胆小管中积累的胆汁酸。我们研究了嵌入不同大小胶原凝胶微腔中的肝细胞中胆小管的形成。使用荧光标记的胆汁酸进行图像分析表明，与其他大小的微腔和使用传统肝细胞三明治培养的肝细胞相比，直径为 60 或 80μm 的微腔中嵌入肝细胞的胆小管面积增大。我们使用玻璃毛细管从微腔中培养的肝细胞的扩大的胆小管中成功回收了胆汁酸，并对回收的量进行了定量。使用我们的方法，通过增强胆汁排泄和几何放大的胆小管的肝细胞培养，可以直接回收胆汁代谢物，从而可以针对药物转运体对药代动力学和药物相互作用进行准确筛选。
