Investigation of calcium mechanotransduction by quasi 3-D microfiber mechanical stimulation of cells

Fibroblast cells mechanically interact with their environment through several mechanisms, including the formation of focal adhesion complexes (FACs) with adjacent ligands. A quasi-3D (Q3D) experimental device was devised to investigate chemo-mechanical stimulation of these cells. It employed a novel microscopic fixture featuring micropolymer arrays positioned just above a cellular monolayer, enabling mechanical stimulation through dorsal attachment to FACs. This protocol demonstrated the ability to preserve the normal stellate morphology of fibroblasts. Live-cell imaging during mechanical stimulation revealed robust calcium elevation in cells interacting with the Q3D device. This system provides a simple alternative to existing 3D culture methods yet allows dynamic, real-time mechanical probing of cells in a physiologically significant environment as compared to optical or magnetic tweezers.