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Novel Method using PEMF Cell Culture Reproducibility

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Publication Title | Novel Method using PEMF Cell Culture Reproducibility

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Brief Report
A Novel Method to Achieve Precision and Reproducibility in Exposure Parameters for Low-Frequency Pulsed Magnetic Fields in Human Cell Cultures
Michael Ronniger 1,*,† , Blanche Aguida 2,† , Christina Stacke 1,† , Yangmengfan Chen 3 , Sabrina Ehnert 3 , Niklas Erdmann 1 , Georg Eschenburg 1 , Karsten Falldorf 1 , Marootpong Pooam 4 , Anthony Wing 1
and Margaret Ahmad 2,*,†
1 2 3 4
* Correspondence: (M.R.); (M.A.); Tel.: +49-408-060-961-25 (M.R.); +33-014-427-2916 (M.A.)
† These authors contributed equally to this work.
Abstract: The effects of extremely low-frequency electromagnetic field (ELF-MF) exposure on living systems have been widely studied at the fundamental level and also claimed as beneficial for the treatment of diseases for over 50 years. However, the underlying mechanisms and cellular targets of ELF-MF exposure remain poorly understood and the field has been plagued with controversy stemming from an endemic lack of reproducibility of published findings. To address this problem, we here demonstrate a technically simple and reproducible EMF exposure protocol to achieve a standardized experimental approach which can be readily adopted in any lab. As an assay system, we chose a commercially available inflammatory model human cell line; its response to magnetic fields involves changes in gene expression which can be monitored by a simple colorimetric reporter gene assay. The cells were seeded and cultured in microplates and inserted into a custom-built, semi- automated incubation and exposure system which accurately controls the incubation (temperature, humidity, CO2) and magnetic-field exposure conditions. A specific alternating magnetic field (<1.0% spatial variance) including far-field reduction provided defined exposure conditions at the position of each well of the microplate. To avoid artifacts, all environmental and magnetic-field exposure parameters were logged in real time throughout the duration of the experiment. Under these extensively controlled conditions, the effect of the magnetic field on the cell cultures as assayed by the standardized operating procedure was highly reproducible between experiments. As we could fully define the characteristics (frequency, intensity, duration) of the pulsed magnetic field signals at the position of the sample well, we were, for the first time, able to accurately determine the effect of changing single ELF-MF parameters such as signal shape, frequency, intensity and duty cycle on the biological response. One signal in particular (10 Hz, 50% duty cycle, rectangular, bipolar, 39.6 μT) provided a significant reduction in cytokine reporter gene expression by 37% in our model cell culture line. In sum, the accuracy, environmental control and data-logging capacity of the semi-automated exposure system should greatly facilitate research into fundamental cellular response mechanisms and achieve the consistency necessary to bring ELF-MF/PEMF research results into the scientific mainstream.
Keywords: HEK-Blueelectromagneticfield;extremelylow-frequency;EMF;PEMF;ELF-MF;inflam- mation; HEK-Blue; TLR4
Sachtleben GmbH, 20251 Hamburg, Germany
Photobiology Research Group, Sorbonne Université CNRS, 75005 Paris, France
Department of Biology, Faculty of Science, Naresuan University, Phitsanulok 65000, Thailand Siegfried Weller Institute for Trauma Research, Department of Trauma and Reconstructive Surgery, BG Unfallklinik Tübingen, Eberhard Karls Universität Tübingen, 72076 Tübingen, Germany
Citation: Ronniger, M.; Aguida, B.; Stacke, C.; Chen, Y.; Ehnert, S.; Erdmann, N.; Eschenburg, G.; Falldorf, K.; Pooam, M.; Wing, A.; et al. A Novel Method to Achieve Precision and Reproducibility in Exposure Parameters for Low-Frequency Pulsed Magnetic Fields in Human Cell Cultures. Bioengineering 2022, 9, 595. bioengineering9100595
Academic Editor: Cornelia Kasper
Received: 29 August 2022 Accepted: 30 September 2022 Published: 21 October 2022
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations.
Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// 4.0/).
Bioengineering 2022, 9, 595.

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