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Application of ECIS to Assess FCCP-Induced Changes of MSC Micromotion and Wound Healing Migration

1
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan
2
Division of Endocrinology and Metabolism, Department of Internal Medicine, Tri-Service General Hospital Songshan Branch, National Defense Medical Center, Taipei 11490, Taiwan
3
Department of Biomedical Engineering, National Yang-Ming University, Taipei 11221, Taiwan
4
Department of Physiology, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan
*
Authors to whom correspondence should be addressed.
Sensors 2019, 19(14), 3210; https://doi.org/10.3390/s19143210
Received: 17 June 2019 / Revised: 19 July 2019 / Accepted: 19 July 2019 / Published: 21 July 2019
(This article belongs to the Section Biosensors)
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Abstract

Electric cell-substrate impedance sensing (ECIS) is an emerging technique for sensitively monitoring morphological changes of adherent cells in tissue culture. In this study, human mesenchymal stem cells (hMSCs) were exposed to different concentrations of carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) for 20 h and their subsequent concentration-dependent responses in micromotion and wound healing migration were measured by ECIS. FCCP disrupts ATP synthesis and results in a decrease in cell migration rates. To detect the change of cell micromotion in response to FCCP challenge, time-series resistances of cell-covered electrodes were monitored and the values of variance were calculated to verify the difference. While Seahorse XF-24 extracellular flux analyzer can detect the effect of FCCP at 3 μM concentration, the variance calculation of the time-series resistances measured at 4 kHz can detect the effect of FCCP at concentrations as low as 1 μM. For wound healing migration, the recovery resistance curves were fitted by sigmoid curve and the hill slope showed a concentration-dependent decline from 0.3 μM to 3 μM, indicating a decrease in cell migration rate. Moreover, dose dependent incline of the inflection points from 0.3 μM to 3 μM FCCP implied the increase of the half time for wound recovery migration. Together, our results demonstrate that partial uncoupling of mitochondrial oxidative phosphorylation reduces micromotion and wound healing migration of hMSCs. The ECIS method used in this study offers a simple and sensitive approach to investigate stem cell migration and its regulation by mitochondrial dynamics. View Full-Text
Keywords: ECIS; human mesenchymal stem cells (hMSCs); mitochondria; carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP); micromotion; wound healing migration ECIS; human mesenchymal stem cells (hMSCs); mitochondria; carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP); micromotion; wound healing migration
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Chiu, S.-P.; Lee, Y.-W.; Wu, L.-Y.; Tung, T.-H.; Gomez, S.; Lo, C.-M.; Wang, J.-Y. Application of ECIS to Assess FCCP-Induced Changes of MSC Micromotion and Wound Healing Migration. Sensors 2019, 19, 3210.

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