Kane Bartholomew J, Zinner Michael J, Yarmush Martin L, Toner Mehmet
Department of Surgery, Brigham and Women's Hospital, 75 Francis Street, Boston, Massachusetts 02115, USA.
Anal Chem. 2006 Jul 1;78(13):4291-8. doi: 10.1021/ac051856v.
Nearly half a billion dollars in resources are lost each time a drug candidate is withdrawn from the market by the Food and Drug Administration (FDA) for reasons of liver toxicity.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 microg/day, sigma = 3.98 microg/day, N = 8) and urea production (109.8 microg/day, sigma = 11.9 microg/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.
每当一种候选药物因肝毒性被美国食品药品监督管理局（FDA）撤出市场时，就会损失近5亿美元的资源。通过使用能够模拟体内肝脏组织对毒性损伤反应的基于肝细胞的系统，有可能减少因肝毒性导致的晚期药物研发失败的数量。在本文中，我们报告了在实现用于高通量肝毒性研究的原代肝细胞阵列这一目标方面所取得的进展。本文描述了一种64（8×8）元件的微流控孔阵列的开发，该阵列能够支持微图案化的原代大鼠肝细胞与3T3-J2成纤维细胞共培养。阵列中的每个孔都以不可寻址的方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，包括使用两个微流控灌注网络为共培养提供独立且持续的细胞培养基和氧气供应。还描述了用于在阵列孔内选择性地对肝细胞和3T3-J2成纤维细胞进行图案化的制造技术。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持续进行稳态白蛋白合成（78.4微克/天，标准差=3.98微克/天，N=8）和尿素生成（109.8微克/天，标准差=11.9微克/天，N=8）。在本文的最后部分，讨论了这些结果与本研究工作的最终目标——开发用于生理相关毒理学研究的原代肝细胞阵列之间的关系。
