High Content Analysis of Human Pluripotent Stem Cell Derived Hepatocytes Reveals Drug Induced Steatosis and Phospholipidosis.
High Content Analysis of Human Pluripotent Stem Cell Derived Hepatocytes Reveals Drug Induced Steatosis and Phospholipidosis.
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DOI：
10.1155/2016/2475631
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发表时间：
2016
影响因子：
4.3
通讯作者：
Edsbagge J
中科院分区：
文献类型：
--
作者：
Pradip A;Steel D;Jacobsson S;Holmgren G;Ingelman-Sundberg M;Sartipy P;Björquist P;Johansson I;Edsbagge J
肝毒性是批准药物撤出市场最常被提及的原因之一。非临床相关的体外和体内测试系统的使用导致高损耗率。将人类诱导多能干细胞 (hiPSC) 分化为表达功能性药物代谢酶的肝细胞样细胞纯培养物的最新进展，为新型、更相关的基于人类细胞的毒性模型开辟了可能性。本研究旨在研究使用 hiPSC 衍生的肝细胞通过基于图像的高内涵分析 (HCA) 进行机械毒性测试。将 hiPSC 衍生的肝细胞暴露于已知通过脂肪变性和磷脂沉积引起肝毒性的药物，测量代表药物引起的肝毒性的不同机制的几个终点。 hiPSC 衍生的肝细胞以 HepG2 细胞系为基准，生成稳健的 HCA 数据，板和批次之间的不精确度较低。在亚细胞毒性浓度下检测测量的不同参数，并根据与先前发表的数据相对应的等摩尔浓度下的损伤程度将化合物分类的顺序（严重、中度、轻度或无毒）。总而言之，本研究展示了如何使用 hiPSC 衍生的肝细胞作为筛选 HCA 药物诱导的肝毒性的平台。
Hepatotoxicity is one of the most cited reasons for withdrawal of approved drugs from the market. The use of nonclinically relevant in vitro and in vivo testing systems contributes to the high attrition rates. Recent advances in differentiating human induced pluripotent stem cells (hiPSCs) into pure cultures of hepatocyte-like cells expressing functional drug metabolizing enzymes open up possibilities for novel, more relevant human cell based toxicity models. The present study aimed to investigate the use of hiPSC derived hepatocytes for conducting mechanistic toxicity testing by image based high content analysis (HCA). The hiPSC derived hepatocytes were exposed to drugs known to cause hepatotoxicity through steatosis and phospholipidosis, measuring several endpoints representing different mechanisms involved in drug induced hepatotoxicity. The hiPSC derived hepatocytes were benchmarked to the HepG2 cell line and generated robust HCA data with low imprecision between plates and batches. The different parameters measured were detected at subcytotoxic concentrations and the order of which the compounds were categorized (as severe, moderate, mild, or nontoxic) based on the degree of injury at isomolar concentration corresponded to previously published data. Taken together, the present study shows how hiPSC derived hepatocytes can be used as a platform for screening drug induced hepatotoxicity by HCA.