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<p>Electrical stimulation remains a promising but insufficiently standardized approach in regenerative medicine, as many<i> in vitro</i> studies lack physically defined stimulation systems and quantitative characterization of cellular electric-field exposure. Here, we present a measurement-based framework integrating real-time electrical recordings, impedance-informed simulations, and electrode characterization under experimental conditions. Human adipose-derived stem cells were stimulated for three days at 20&#xa0;Hz using sinusoidal alternating current at applied voltages of 1–3 Vrms. Simulations revealed spatially heterogeneous electric-field distributions, with exposure predominantly below 5 Vm⁻¹ at 1 Vrms and shifting toward 5–15 Vm⁻¹ at higher amplitudes. Moderate stimulation (1–2 Vrms) was not associated with detectable reductions in metabolic activity and was accompanied by maintained functional behaviour and stable reference gene expression, whereas 3 Vrms coincided with electrochemical interface alterations and reduced gene stability, suggesting the onset of stress-associated cellular responses. These findings identify a system-specific, biologically compatible and functionally active stimulation window and demonstrate that cellular responses are governed by the effective electrical dose rather than nominal voltage alone. The presented framework enables reproducible, impedance-controlled and quantitatively defined <i>in vitro</i> alternating-current stimulation.</p>

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A validated in vitro approach for low-frequency alternating-current stimulation of stem cells: From electrode characterization to biological responses

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月18日

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  • Laura Lembcke,
  • Henning Bathel,
  • Julius Zimmermann,
  • Vivien Engel,
  • Rainer Bader,
  • Bernhard Frerich,
  • Ursula van Rienen,
  • Armin Springer,
  • Nadja Engel,
  • Peer W. Kämmerer

摘要

Electrical stimulation remains a promising but insufficiently standardized approach in regenerative medicine, as many in vitro studies lack physically defined stimulation systems and quantitative characterization of cellular electric-field exposure. Here, we present a measurement-based framework integrating real-time electrical recordings, impedance-informed simulations, and electrode characterization under experimental conditions. Human adipose-derived stem cells were stimulated for three days at 20 Hz using sinusoidal alternating current at applied voltages of 1–3 Vrms. Simulations revealed spatially heterogeneous electric-field distributions, with exposure predominantly below 5 Vm⁻¹ at 1 Vrms and shifting toward 5–15 Vm⁻¹ at higher amplitudes. Moderate stimulation (1–2 Vrms) was not associated with detectable reductions in metabolic activity and was accompanied by maintained functional behaviour and stable reference gene expression, whereas 3 Vrms coincided with electrochemical interface alterations and reduced gene stability, suggesting the onset of stress-associated cellular responses. These findings identify a system-specific, biologically compatible and functionally active stimulation window and demonstrate that cellular responses are governed by the effective electrical dose rather than nominal voltage alone. The presented framework enables reproducible, impedance-controlled and quantitatively defined in vitro alternating-current stimulation.