Enhanced bile canaliculi formation enabling direct recovery of biliary metabolites of hepatocytes in 3D collagen gel microcavities
Enhanced bile canaliculi formation enabling direct recovery of biliary metabolites of hepatocytes in 3D collagen gel microcavities
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DOI：
10.1039/c2lc40046d
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发表时间：
2012-01-01
期刊：
影响因子：
6.1
通讯作者：
Sakai, Yasuyuki
中科院分区：
文献类型：
--
作者：
Matsui, Hitoshi;Takeuchi, Shoji;Sakai, Yasuyuki
在药物开发过程中，胆道代谢物的分析对于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配置，通过将肝细胞包埋在3D微图案胶原凝胶底物中，可以直接恢复胆管中积累的胆汁酸。我们研究了肝细胞包埋在不同大小的由胶原凝胶制成的圆形微腔中的胆管的形成。荧光标记胆汁酸图像分析显示，在直径为60或80 μ m的微腔中，与其他大小的微腔和传统肝细胞夹层培养的肝细胞相比，包埋肝细胞的胆管面积增大。我们成功地利用玻璃毛细管从微腔培养的肝细胞的胆管中回收胆汁酸，并定量回收量。使用我们的方法，直接恢复胆道代谢物，使用肝细胞培养增强胆道排泄和几何形状扩大的胆管，可以准确筛选药物代动力学和药物-药物相互作用对抗药物转运体。
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 mu m 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.