Integrated Workflow for Electromechanical Stimulation of Cells in Active Scaffolds: From Live Cell Imaging to Functional Assays

In vivo, cells are exposed to a dynamic extracellular matrix (ECM) that delivers biochemical, biophysical, and electrical cues essential for regulating cellular behavior. However, most existing in vitro culture systems remain static and fail to reproduce the time-dependent mechanical and electrical signals that characterize native tissues. Developing biomimetic and dynamic platforms that bridge this gap is therefore critical for advancing mechanobiology, tissue engineering, or drug screening studies. Here, we report a 4D cell culture platform based on polyHIPE-PEDOT scaffolds, combining the structural versatility of polyHIPE architectures synthesized from high internal phase emulsion with the electroactive behavior of the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT). The resulting electroactive scaffolds exhibit a highly interconnected porous structure with tunable morphology and demonstrate reversible pore deformation under electrical stimulation. To facilitate a broad range of experimental applications, we designed two complementary devices: one optimized for in vitro cell culture under standard incubator conditions, and another tailored for real-time live-cell imaging. To illustrate this integrated workflow, we provide examples with fibroblast cells cultured under electromechanical stimulation using our two devices. The objective of this workflow is to facilitate novel insights into the study of time-resolved cell-matrix interactions and to develop responsive biomimetic microenvironments for advanced biomedical research.