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The culture of anchorage-dependent cells is crucial in the biomedical industry, yet traditional dissociation methods, including enzymatic and mechanical techniques, often reduce cell viability and induce cellular stress, particularly in sensitive primary cell populations. These approaches are also resource-intensive, generate considerable biological waste, and lack compatibility with scalable or automated platforms. Here, we propose an enzyme-free and on-demand cell detachment strategy utilizing alternating electrochemical redox-cycling. This approach induces reversible morphological changes that promote cell detachment while maintaining high viability and stable proliferation. When applied to MG63 human osteosarcoma cells on a poly(3,4-ethylenedioxythiophene) polystyrenesulfonate nanocomposite biointerface, the application of voltage initiates redox-cycling, generating ion flux that disrupts cell adhesion and facilitates rounding within 5 min. Detachment efficiency increases from 1 to 95% at an optimal frequency of 0.05 Hz, with cell viability exceeding 90%, demonstrating the feasibility and effectiveness of this method. We introduce an efficient and enzyme-free solution for cell harvesting, which is compatible with automated cell culture and biomanufacturing workflows.