Cell line-based liver models are critical tools for liver-related studies. However, the conventional monolayer culture of hepatocytes, the most widely used in vitro model, does not have the extracellular matrix (ECM), which contributes to the three-dimensional (3D) arrangement of the hepatocytes in the liver. As a result, the metabolic properties of the hepatocytes in the monolayer tissue culture may not accurately reflect those of the hepatocytes in the liver. Here, we developed a modular platform for 3D hepatocyte cultures on fibrous ECMs produced by electrospinning, a technique that can turn a polymer solution to the micro/nanofibers and has been widely used to produce scaffolds for 3D cell cultures. Metabolomics quantitation by liquid chromatography–mass spectrometry (LC–MS) indicated that Huh7 hepatocytes grown in microfibers electrospun from silk fibroin exhibited reduced glycolysis and tricarboxylic acid (TCA) cycle, as compared to the cells cultured as a monolayer. Further mechanistic studies suggested that integrins were correlated to the ECM’s effects. This is the first time to report how an ECM scaffold could affect the fundamental metabolism of the hepatocytes via integrins.

中文翻译：

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微纤维细胞外基质通过整合素改变肝细胞能量代谢

基于细胞系的肝脏模型是进行肝脏相关研究的重要工具。然而，肝细胞的常规单层培养是体外使用最广泛的该模型没有细胞外基质（ECM），而该基质有助于肝脏中肝细胞的三维（3D）排列。结果，单层组织培养物中肝细胞的代谢特性可能无法准确反映肝脏中肝细胞的代谢特性。在这里，我们为通过静电纺丝生产的纤维ECM上的3D肝细胞培养开发了一个模块化平台，该技术可以将聚合物溶液转变为微纤维/纳米纤维，并且已广泛用于生产3D细胞培养的支架。通过液相色谱-质谱（LC-MS）进行的代谢组学定量分析表明，与以单层培养的细胞相比，用丝素蛋白静电纺丝的超细纤维中生长的Huh7肝细胞显示出降低的糖酵解和三羧酸（TCA）循环。进一步的机理研究表明，整联蛋白与ECM的作用有关。这是首次报道ECM支架如何通过整联蛋白影响肝细胞的基本代谢。