Human kidney proximal tubule-on-a-chip for drug transport and nephrotoxicity assessment
Human kidney proximal tubule-on-a-chip for drug transport and nephrotoxicity assessment
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DOI：
10.1039/c3ib40049b
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发表时间：
2013-01-01
影响因子：
2.5
通讯作者：
Ingber, Donald E.
中科院分区：
文献类型：
--
作者：
Jang, Kyung-Jin;Mehr, Ali Poyan;Ingber, Donald E.
肾毒性是药物开发过程中最常见的不良事件之一。由于缺乏准确的预测细胞培养模型和动物研究的不可靠性，需要更好的方法来重现体外肾功能。在这里，我们描述了一个微流体装置内衬活的人肾上皮细胞暴露于流体流，模仿人类肾脏近端小管的关键功能。从人近端小管分离的原代肾上皮细胞在细胞外基质包被的多孔聚酯膜的上表面上培养，该膜将装置的主通道分成两个相邻通道，从而产生顶端“管腔”通道和基底“间质”空间。与传统的Transwell培养系统相比，将上皮单层暴露于模拟在活肾小管中发现的顶端流体剪切应力（0.2达因cm（-2））导致增强的上皮细胞极化和初级纤毛形成。细胞还表现出显着更大的白蛋白转运，葡萄糖重吸收，刷状缘碱性磷酸酶活性。重要的是，顺铂毒性和Pgp外排转运蛋白活性的芯片上测量更接近模拟体内反应比在常规培养条件下保持细胞获得的结果。虽然过去的研究已经分析了在体外流动条件下培养的肾小管细胞，但这是第一次报告在微流体“器官芯片”微装置中使用原代人肾近端肾小管上皮细胞进行毒性研究。在该系统中观察到的体内样病理生理学表明，其可作为临床前安全性研究中评价人类相关肾毒性的有用工具。
Kidney toxicity is one of the most frequent adverse events reported during drug development. The lack of accurate predictive cell culture models and the unreliability of animal studies have created a need for better approaches to recapitulate kidney function in vitro. Here, we describe a microfluidic device lined by living human kidney epithelial cells exposed to fluidic flow that mimics key functions of the human kidney proximal tubule. Primary kidney epithelial cells isolated from human proximal tubule are cultured on the upper surface of an extracellular matrix-coated, porous, polyester membrane that splits the main channel of the device into two adjacent channels, thereby creating an apical 'luminal' channel and a basal 'interstitial' space. Exposure of the epithelial monolayer to an apical fluid shear stress (0.2 dyne cm(-2)) that mimics that found in living kidney tubules results in enhanced epithelial cell polarization and primary cilia formation compared to traditional Transwell culture systems. The cells also exhibited significantly greater albumin transport, glucose reabsorption, and brush border alkaline phosphatase activity. Importantly, cisplatin toxicity and Pgp efflux transporter activity measured on-chip more closely mimic the in vivo responses than results obtained with cells maintained under conventional culture conditions. While past studies have analyzed kidney tubular cells cultured under flow conditions in vitro, this is the first report of a toxicity study using primary human kidney proximal tubular epithelial cells in a microfluidic 'organ-on-a-chip' microdevice. The in vivo-like pathophysiology observed in this system suggests that it might serve as a useful tool for evaluating human-relevant renal toxicity in preclinical safety studies.