Chang Shih-Yu, Voellinger Jenna L, Van Ness Kirk P, Chapron Brian, Shaffer Rachel M, Neumann Thomas, White Collin C, Kavanagh Terrance J, Kelly Edward J, Eaton David L
Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195, USA.
Department of Pharmaceutics, University of Washington, Seattle, WA 98195, USA.
Toxicol In Vitro. 2017 Apr;40:170-183. doi: 10.1016/j.tiv.2017.01.007. Epub 2017 Jan 13.
The liver is the main site for drug and xenobiotics metabolism, including inactivation or bioactivation. In order to improve the predictability of drug safety and efficacy in clinical development, and to facilitate the evaluation of the potential human health effects from exposure to environmental contaminants, there is a critical need to accurately model human organ systems such as the liver in vitro. We are developing a microphysiological system (MPS) based on a new commercial microfluidic platform (Nortis, Inc.) that can utilize primary liver cells from multiple species (e.g., rat and human). Compared to conventional monolayer cell culture, which typically survives for 5-7days or less, primary rat or human hepatocytes in an MPS exhibited higher viability and improved hepatic functions, such as albumin production, expression of hepatocyte marker HNF4α and canaliculi structure, for up to 14days. Additionally, induction of Cytochrome P450 (CYP) 1A and 3A4 in cryopreserved human hepatocytes was observed in the MPS. The acute cytotoxicity of the potent hepatotoxic and hepatocarcinogen, aflatoxin B, was evaluated in human hepatocytes cultured in an MPS, demonstrating the utility of this model for acute hepatotoxicity assessment. These results indicate that MPS-cultured hepatocytes provide a promising approach for evaluating chemical toxicity in vitro.
肝脏是药物和异生素代谢的主要场所，包括失活或生物活化。为了提高临床开发中药物安全性和有效性的可预测性，并促进对接触环境污染物可能对人类健康产生的影响进行评估，迫切需要在体外准确模拟人体器官系统，如肝脏。我们正在基于一种新的商业微流控平台（Nortis公司）开发一种微生理系统（MPS），该平台可以利用多种物种（如大鼠和人类）的原代肝细胞。与通常存活5-7天或更短时间的传统单层细胞培养相比，MPS中的原代大鼠或人类肝细胞表现出更高的活力和改善的肝功能，如白蛋白产生、肝细胞标志物HNF4α的表达和胆小管结构，长达14天。此外，在MPS中观察到冷冻保存的人肝细胞中细胞色素P450（CYP）1A和3A4的诱导。在MPS中培养的人肝细胞中评估了强效肝毒性和肝癌致癌物黄曲霉毒素B的急性细胞毒性，证明了该模型在急性肝毒性评估中的实用性。这些结果表明，MPS培养的肝细胞为体外评估化学毒性提供了一种有前景的方法。
