PDF(Pubmed)
Abstract：
Drug-induced cholestasis results in drug discontinuation and market withdrawal, and the prediction of cholestasis risk is critical in the early stages of drug development. Animal tests and membrane vesicle assay are currently being conducted to assess the risk of cholestasis in the preclinical stage. However, these methods have drawbacks, such as species differences with humans and difficulties in evaluating the effects of drug metabolism and other transporters, implying the need for a cholestasis risk assessment system using human hepatocytes. However, human hepatocytes hardly form functional, extended bile canaliculi, a requirement for cholestasis risk assessment. We previously established a culture protocol for functional, extended bile canaliculi formation in human iPSC-derived hepatocytes. In this study, we modified this culture protocol to support the formation of functional, extended bile canaliculi in human cryopreserved hepatocytes (cryoheps). The production of bile acids, which induces bile canaliculi extension, increased time-dependently during bile canaliculi formation using this protocol, suggesting that increased bile acid production may be involved in the extended bile canaliculi formation. We have also shown that our culture protocol can be applied to cryoheps from multiple donors and that bile canaliculi can be formed stably among different culture batches. Furthermore, this protocol enables long-term maintenance of bile canaliculi and scaling down to culture in 96-well plates. We expect our culture protocol to be a breakthrough for in vitro cholestasis risk assessment.
摘要：
药物引起的胆汁淤积导致药物停药和市场退出，在药物开发的早期阶段，对胆汁淤积风险的预测至关重要。目前正在进行动物测试和膜囊泡测定，以评估临床前阶段胆汁淤积的风险。然而,这些方法有缺点，例如与人类的物种差异以及评估药物代谢和其他转运蛋白的影响的困难，这意味着需要使用人肝细胞的胆汁淤积风险评估系统。然而,人肝细胞几乎没有形成功能，延伸的胆管，胆汁淤积风险评估的要求。我们之前建立了一个培养协议的功能，在人iPSC衍生的肝细胞中延长的胆小管形成。在这项研究中,我们修改了这个文化协议来支持功能的形成，在人冷冻保存的肝细胞（冷冻肝）中延长的胆管。胆汁酸的产生，诱导胆管扩张，使用该方案在胆小管形成过程中时间依赖性增加，提示胆汁酸的产生增加可能与胆小管的形成有关。我们还表明，我们的培养方案可以应用于来自多个供体的冷冻肠，并且可以在不同培养批次之间稳定地形成胆管。此外，该方案能够长期维持胆管和缩小到96孔板中的培养。我们预计我们的培养方案将成为体外胆汁淤积风险评估的突破。
