Wu Yun-Chen, Yang Jing-Yi, Hsu Chia-Hsien
Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan, Miaoli 35053, Taiwan.
Institute of Nanoengineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan.
R Soc Open Sci. 2024 Aug 14;11(8):240284. doi: 10.1098/rsos.240284. eCollection 2024 Aug.
Advanced culture systems have emerged as alternatives to animal testing and traditional cell culture methods in biomedical research. Polydimethylsiloxane (PDMS) is frequently used in creating sophisticated culture devices owing to its elastomeric properties, which allow mechanical stretching to simulate physiological movements in cell experiments. We introduce a straightforward method that uses three types of commercial tape-generic, magic and masking-to fabricate PDMS membranes with microscale thicknesses (47.2 µm for generic, 58.1 µm for magic and 89.37 µm for masking) in these devices. These membranes are shaped as the bases of culture wells and can perform cyclic radial movements controlled via a vacuum system. In experiments with A549 cells under three mechanical stimulation conditions, we analysed transcriptional regulators responsive to external mechanical stimuli. Results indicated increased nuclear yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) activity in both confluent and densely packed cells under cyclically mechanical strains (Pearson's coefficient (PC) of 0.59 in confluent and 0.24 in dense cells) compared with static (PC = 0.47 in confluent and 0.13 in dense) and stretched conditions (PC = 0.55 in confluent and 0.20 in dense). This technique offers laboratories without microfabrication capabilities a viable option for exploring cellular behaviour under dynamic mechanical stimulation using PDMS membrane-equipped devices.
在生物医学研究中，先进的培养系统已成为动物实验和传统细胞培养方法的替代方案。聚二甲基硅氧烷（PDMS）因其弹性特性，常被用于制造精密的培养装置，这种特性使得在细胞实验中能够通过机械拉伸来模拟生理运动。我们介绍一种简单的方法，该方法使用三种商用胶带——普通胶带、魔术胶带和遮蔽胶带，在这些装置中制造具有微米级厚度（普通胶带为47.2 µm，魔术胶带为58.1 µm，遮蔽胶带为89.37 µm）的PDMS膜。这些膜被制成培养孔的底部形状，并能通过真空系统控制进行周期性的径向运动。在对A549细胞进行的三种机械刺激条件下的实验中，我们分析了对外部机械刺激有反应的转录调节因子。结果表明，与静态（汇合细胞中皮尔逊系数（PC）为0.47，密集细胞中为0.13）和拉伸条件（汇合细胞中PC为0.55，密集细胞中为0.20）相比，在周期性机械应变下，汇合和密集细胞中的核Yes相关蛋白（YAP）和含PDZ结合基序的转录共激活因子（TAZ）活性均增加（汇合细胞中PC为0.59，密集细胞中为0.24）。这项技术为没有微加工能力的实验室提供了一种可行的选择，可使用配备PDMS膜的装置来探索动态机械刺激下的细胞行为。
