Mimicking liver sinusoidal structures and functions using a 3D-configured microfluidic chip
Mimicking liver sinusoidal structures and functions using a 3D-configured microfluidic chip
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使用 3D 配置的微流控芯片模拟肝窦结构和功能
DOI：
10.1039/c6lc01374k
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发表时间：
2017-03-07
期刊：
影响因子：
6.1
通讯作者：
Long, Mian
中科院分区：
文献类型：
--
作者：
Du, Yu;Li, Ning;Long, Mian
在生理学上，四种主要类型的肝细胞-肝窦内皮细胞、枯否细胞、肝星状细胞和肝细胞-驻留在肝窦内并在血流下与流动的外周细胞相互作用。由于肝窦复杂的细胞间相互作用、时空结构和力学微环境，很难在体内模拟肝窦。在这里，我们开发了一种体外肝窦状隙芯片，通过将四种类型的原代小鼠肝细胞整合到由多孔渗透膜分隔的两个相邻的流体通道中，复制肝脏的关键结构和配置。每种类型的细胞用其各自的标记物鉴定，并且组装的芯片呈现肝脏特异性的开窗的独特形态。通过计算流体力学模拟和粒子跟踪可视化实验对肝芯片内部流场进行了定量分析。有趣的是，共培养和剪切流独立或协同地增强白蛋白分泌，而单独的剪切流增强HGF产生和CYP 450代谢。在脂多糖（LPS）刺激下，肝细胞共培养促进了肝芯片中的中性粒细胞募集。因此，这种3D配置的体外肝脏芯片集成了剪切流的两个关键因素和四种类型的原代肝细胞，以复制关键结构，肝功能和初级免疫反应，并提供了一种新的体外模型，以研究在模拟肝脏生理学的微环境下的短期肝细胞相互作用。
Physiologically, four major types of hepatic cells - the liver sinusoidal endothelial cells, Kupffer cells, hepatic stellate cells, and hepatocytes - reside inside liver sinusoids and interact with flowing peripheral cells under blood flow. It is hard to mimic an in vivo liver sinusoid due to its complex multiple cell-cell interactions, spatiotemporal construction, and mechanical microenvironment. Here we developed an in vitro liver sinusoid chip by integrating the four types of primary murine hepatic cells into two adjacent fluid channels separated by a porous permeable membrane, replicating liver's key structures and configurations. Each type of cells was identified with its respective markers, and the assembled chip presented the liver-specific unique morphology of fenestration. The flow field in the liver chip was quantitatively analyzed by computational fluid dynamics simulations and particle tracking visualization tests. Intriguingly, co-culture and shear flow enhance albumin secretion independently or cooperatively, while shear flow alone enhances HGF production and CYP450 metabolism. Under lipopolysaccharide (LPS) stimulations, the hepatic cell co-culture facilitated neutrophil recruitment in the liver chip. Thus, this 3D-configured in vitro liver chip integrates the two key factors of shear flow and the four types of primary hepatic cells to replicate key structures, hepatic functions, and primary immune responses and provides a new in vitro model to investigate the short-duration hepatic cellular interactions under a microenvironment mimicking the physiology of a liver.