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.