Tape-assisted fabrication method for constructing PDMS membrane-containing culture devices with cyclic radial stretching stimulation

Advanced in vitro 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 due to its elastomeric properties, which allow mechanical stretching to simulate physiological movements in cell experiments. We introduce a straightforward method that utilizes 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 analyzed transcriptional regulators responsive to external mechanical stimuli. Results indicated increased nuclear YAP/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 to 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 behavior under dynamic mechanical stimulation using PDMS membrane-equipped devices.