School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China.
The CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China.
Int J Mol Sci. 2024 Jan 16;25(2):1081. doi: 10.3390/ijms25021081.
Three-dimensional (3D) hepatocyte models have become a research hotspot for evaluating drug metabolism and hepatotoxicity. Compared to two-dimensional (2D) cultures, 3D cultures are better at mimicking the morphology and microenvironment of hepatocytes in vivo. However, commonly used 3D culture techniques are not suitable for high-throughput drug screening (HTS) due to their high cost, complex handling, and inability to simulate cell-extracellular matrix (ECM) interactions. This article describes a method for rapid and reproducible 3D cell cultures with ECM-cell interactions based on 3D culture instrumentation to provide more efficient HTS. We developed a microsphere preparation based on a high-voltage electrostatic (HVE) field and used sodium alginate- and collagen-based hydrogels as scaffolds for 3D cultures of HepG2 cells. The microsphere-generating device enables the rapid and reproducible preparation of bioactive hydrogel microspheres. This 3D culture system exhibited better cell viability, heterogeneity, and drug-metabolizing activity than 2D and other 3D culture models, and the long-term culture characteristics of this system make it suitable for predicting long-term liver toxicity. This system improves the overall applicability of HepG2 spheroids in safety assessment studies, and this simple and controllable high-throughput-compatible method shows potential for use in drug toxicity screening assays and mechanistic studies.
三维（3D）肝细胞模型已成为评估药物代谢和肝毒性的研究热点。与二维（2D）培养相比，3D 培养更能模拟肝细胞在体内的形态和微环境。然而，常用的 3D 培养技术由于成本高、操作复杂以及无法模拟细胞-细胞外基质（ECM）相互作用，不适合高通量药物筛选（HTS）。本文描述了一种基于 3D 培养仪器的具有 ECM-细胞相互作用的快速、可重复的 3D 细胞培养方法，以提供更有效的 HTS。我们开发了一种基于高压静电（HVE）场的微球制备方法，并使用海藻酸钠和胶原蛋白基水凝胶作为 HepG2 细胞 3D 培养的支架。微球生成装置能够快速、可重复地制备生物活性水凝胶微球。与 2D 和其他 3D 培养模型相比，该 3D 培养系统表现出更好的细胞活力、异质性和药物代谢活性，并且该系统的长期培养特性使其适合预测长期肝毒性。该系统提高了 HepG2 球体在安全性评估研究中的整体适用性，这种简单可控的高通量兼容方法在药物毒性筛选测定和机制研究中具有应用潜力。
