Liver-specific functional studies in a microfluidic array of primary mammalian hepatocytes
Liver-specific functional studies in a microfluidic array of primary mammalian hepatocytes
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DOI：
10.1021/ac051856v
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发表时间：
2006-07-01
影响因子：
7.4
通讯作者：
Toner, Mehmet
中科院分区：
文献类型：
--
作者：
Kane, Bartholomew J.;Zinner, Michael J.;Toner, Mehmet
每次美国食品和药物管理局 (FDA) 因肝毒性原因将候选药物从市场上撤回时，都会损失近 5 亿美元的资源。通过使用能够模拟体内肝组织对毒性损伤反应的基于肝细胞的系统，可能会减少由于肝毒性而导致的后期药物开发失败的数量。在本文中，我们报告了实现一系列原代肝细胞用于高通量肝毒性研究的目标的进展。本文描述的是 64 (8 x 8) 元件微流体孔阵列的开发，该阵列能够支持与 3T3-J2 成纤维细胞共培养的微图案原代大鼠肝细胞。阵列内的每个孔以不可寻址的形式连续灌注培养基和氧气。描述了系统设计和制造的关键特征，包括使用两个微流体灌注网络为共培养物提供独立且连续的细胞培养基和氧气供应。还描述了用于选择性地图案化阵列孔内的肝细胞和 3T3-J2 成纤维细胞的制造技术。本文概述了用于证明该阵列的合成和代谢能力的功能研究。这些研究表明，阵列中包含的肝细胞能够连续、稳态地合成白蛋白（78.4μg/天，sigma = 3.98μg/天，N = 8）和尿素生产（109.8μg/天，sigma = 11.9μg/天，N）8）。在本文的最后部分，讨论了这些结果，因为它们与本研究工作的最终目标相关，即开发一系列原代肝细胞用于生理相关毒理学研究。
Nearly half a billion dollars in resources are lost each time a drug candidate is withdrawn from the market by the Food and Drug Administration (FDA) for reasons of liver toxicity. The number of late-phase drug developmental failures due to liver toxicity could potentially be reduced through the use of hepatocyte-based systems capable of modeling the response of in vivo liver tissue to toxic insults. With this article, we report progress toward the goal of realizing an array of primary hepatocytes for use in high-throughput liver toxicity studies. Described herein is the development of a 64 ( 8 x 8) element array of microfluidic wells capable of supporting micropatterned primary rat hepatocytes in coculture with 3T3-J2 fibroblasts. Each of the wells within the array was continuously perfused with medium and oxygen in a nonaddressable format. The key features of the system design and fabrication are described, including the use of two microfluidic perfusion networks to provide the coculture with an independent and continuous supply of cell culture medium and oxygen. Also described are the fabrication techniques used to selectively pattern hepatocytes and 3T3-J2 fibroblasts within the wells of the array. The functional studies used to demonstrate the synthetic and metabolic capacity of the array are outlined in this article. These studies demonstrate that the hepatocytes contained within the array are capable of continuous, steady-state albumin synthesis (78.4 mu g/day, sigma = 3.98 mu g/ day, N = 8) and urea production (109.8 mu g/day, sigma = 11.9 mu g/day, N) 8). In the final section of the article, these results are discussed as they relate to the final goal of this research effort, the development of an array of primary hepatocytes for use in physiologically relevant toxicology studies.