用于生理学相关病毒感染研究的离心微流控细胞培养平台:HSV-1 感染牙周细胞的案例研究。

IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL
Juliane Fjelrad Christfort, Morgane Ortis, Hau Van Nguyen, Robert Marsault, Alain Doglio
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引用次数: 0

摘要

静态孔板仍然是研究体外病毒感染的黄金标准,但它无法准确模拟人体内发生的动态病毒感染。因此,我们建立了一个基于离心微流控技术的动态细胞培养平台,以研究灌流中的病毒感染。为此,我们以人类牙周韧带(PDL)原代细胞和单纯疱疹病毒-1(HSV-1)为案例进行了研究。通过显微镜观察,我们证实 PDL 细胞能在芯片中有效附着和生长。通过荧光成像和基于 RT-qPCR 的实验监测了灌注 PDL 细胞成功的动态病毒感染。值得注意的是,流式病毒感染导致 HSV-1 感染细胞的梯度从细胞培养室入口向末端逐渐降低。在芯片中灌注阿昔洛韦阻止了 HSV-1 的扩散,证明了这种平台在监测抗病毒药物效果方面的实用性。此外,与灌流模型相比,通过干扰素基因表达衡量的 PDL 细胞先天抗病毒反应在传统静态条件下随着时间的推移显著增强。这些结果提供的证据表明,动态病毒感染不同于传统的静态感染,这突出表明需要更多与生理相关的体外模型来研究病毒感染。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Centrifugal Microfluidic Cell Culture Platform for Physiologically Relevant Virus Infection Studies: A Case Study with HSV-1 Infection of Periodontal Cells.

Static well plates remain the gold standard to study viral infections in vitro, but they cannot accurately mimic dynamic viral infections as they occur in the human body. Therefore, we established a dynamic cell culture platform, based on centrifugal microfluidics, to study viral infections in perfusion. To do so, we used human primary periodontal dental ligament (PDL) cells and herpes simplex virus-1 (HSV-1) as a case study. By microscopy, we confirmed that the PDL cells efficiently attached and grew in the chip. Successful dynamic viral infection of perfused PDL cells was monitored using fluorescent imaging and RT-qPCR-based experiments. Remarkably, viral infection in flow resulted in a gradient of HSV-1-infected cells gradually decreasing from the cell culture chamber entrance towards its end. The perfusion of acyclovir in the chip prevented HSV-1 spreading, demonstrating the usefulness of such a platform for monitoring the effects of antiviral drugs. In addition, the innate antiviral response of PDL cells, measured by interferon gene expression, increased significantly over time in conventional static conditions compared to the perfusion model. These results provide evidence suggesting that dynamic viral infections differ from conventional static infections, which highlights the need for more physiologically relevant in vitro models to study viral infections.

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来源期刊
Biosensors-Basel
Biosensors-Basel Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
6.60
自引率
14.80%
发文量
983
审稿时长
11 weeks
期刊介绍: Biosensors (ISSN 2079-6374) provides an advanced forum for studies related to the science and technology of biosensors and biosensing. It publishes original research papers, comprehensive reviews and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.
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