Developing a Flow-Resistance Module for Elucidating Cell Mechanotransduction on Multiple Shear Stresses

Fluid shear stress plays a pivotal role in regulating cellular behaviors, maintaining tissue homeostasis, and driving disease progression. Cells in various tissues are specifically adapted to physiological levels of shear stress and exhibit sensitivity to variations in its magnitude, highlighting the requirement for a comprehensive understanding of cellular responses to both physiologically and pathologically relevant levels of shear stress. In this study, we developed an independent upstream flow-resistance module with high fluidic resistances comprising three microchannels. The validity of the flow-resistance module was confirmed via computational fluid dynamics (CFD) simulations and flow calibration experiments, resulting in the generation of steady wall shear stresses ranging from 0.06 to 11.57 dyn/cm² within the interconnected cell culture chips. Gene expression profiles, cytoskeletal remodeling, and morphological changes, as well as Yes-associated protein (YAP) nuclear translocation, were investigated in response to various shear stresses to authenticate the reliability of our experimental platform, indicating an increasing trend as the shear stress increases, reaching its maximum at various shear stresses. Our findings suggest that this flow-resistance module can be readily employed for precise characterization of cellular responses under various shear stresses.