Research Article

Visualizing Age-Dependent Electrotaxis of Human Adipose-Derived Stem Cells Under Direct Current Electric Fields

DOI:

10.3791/69666

⸱

December 19th, 2025

 ,  ,  ,  , 
1Department of Plastic Surgery, The First Affiliated Hospital of China Medical University, 2Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, 3Department of Ophthalmology and Vision Science, Institute for Regenerative Cures, University of California, Davis

In This Article

Summary

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In a small cohort of female donors, aging reduced the electrotactic migration of hADSCs. DCEF altered gene expression (747 up, 624 down), enriching sodium transport/channel GO terms and PI3K-Akt KEGG pathways, suggesting involvement in age-related electrotaxis, without establishing causality.

Abstract

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Human adipose-derived stem cells (hADSCs) are pivotal for tissue regeneration and wound healing, and their directed migration is a prerequisite for exerting these therapeutic effects. Electric fields (EFs) are well-recognized as key cues guiding cell migration during wound repair, yet the electrotactic behavior of hADSCs -- especially how donor characteristics (e.g., age) regulate this behavior and its underlying molecular mechanisms -- remains poorly understood. This knowledge gap limits the optimized application of hADSCs in regenerative medicine. In this study, we first validated the existence of electrotaxis in hADSCs and confirmed its voltage dependence: under direct current electric fields (DCEFs) of 100-200 mV/mm, hADSCs migrated directionally toward the anode, with stronger EF intensities enhancing both migration directionality and speed (versus at 0 mV/mm control). We then compared hADSCs from young female donors (27.00 ± 4.58 years) and elderly female donors (62.33 ± 4.04 years) using RNA sequencing (RNA-seq) after 200 mV/mm DCEF stimulation. Transcriptomic analysis identified 747 upregulated and 624 downregulated genes in elderly hADSCs, with differentially expressed genes (DEGs) enriched in biological processes/pathways critical for electrotaxis -- including sodium ion transmembrane transport, voltage-gated sodium channel activity (GO terms), and the PI3K-Akt signaling pathway (KEGG pathway). Functionally, elderly hADSCs exhibited significantly reduced anodal migration (decreased accumulated distance, Euclidean distance, and directness) compared to young hADSCs under DCEF stimulation. This study provides the first evidence of age-dependent electrotaxis in hADSCs, demonstrating that donor age correlates with impaired electrotactic capacity. It further reveals that dysregulated sodium channel activity and PI3K-Akt signaling may underlie this age-related decline. These findings point to the potential regulatory mechanisms of hADSC electrotaxis and offer new insights for tailoring hADSC-based therapies (e.g., selecting optimal donors or targeting sodium/PI3K-Akt pathways) to improve tissue regeneration and wound healing outcomes.

Introduction

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Adipose-derived stem cells (ADSC) possess strong self-renewal capacity and significant regenerative potential. Under appropriate induction conditions, they can differentiate into osteoblasts, chondrocytes, and adipocytes, among others1. Through paracrine signaling, ADSC can also support the generation of blood vessels and lymphatic vessels2,3, inhibit scar formation4, and exhibit immune regulatory effects5. Therefore, ADSC is considered an ideal seed cell for cell-based therapies in regenerative medicine.

The fu....

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Protocol

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Informed consent was obtained from all participants prior to tissue collection. This study was approved by the Ethics Committee of the First Affiliated Hospital of China Medical University (Approval No.: [2018]2018-110-2). Written informed consent was obtained from all participants prior to tissue collection

hADSC isolation and culture
Tissue collection: Human abdominal adipose tissue samples were obtained during routine elective liposuction procedures performed at the Department of Plastic Surgery of the First Affiliated Hospital of China Medical University. All donors were healthy female volunteers ....

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Results

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Application of electric fields guides the directional migration of hADSCs and enhances their migratory capacity
The extracted hADSCs exhibited typical mesenchymal stem cell surface markers (e.g., CD29, CD44, CD90, CD73, CD105) and possessed trilineage differentiation potential (adipogenic, osteogenic, chondrogenic; Figure 2A), meeting the standard criteria for hADSC identification. We applied direct current electric fields (DCEFs) at intensities of 0 mV/mm, 100 mV/mm, .......

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Discussion

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Early in the tissue injury repair and regeneration process, electric fields emerge as one of the earliest physical cues established in the microenvironment. Their directional properties are spatiotemporally correlated with the recruitment of cells to the injury site27. Endogenous electric fields originate from the transepithelial potential generated by polarized ion transport in epithelial tissues. When the epithelial barrier is disrupted or undergoes aging, the potential is secondarily altered, w.......

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Disclosures

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The authors have no conflicts of interest to disclose.

Acknowledgements

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We sincerely acknowledge the Department of Plastic Surgery at The First Affiliated Hospital of China Medical University (Shenyang, China) and the Institute for Regenerative Cures at the University of California, Davis (CA, USA) for their provision of critical experimental infrastructure and shared research resources that enabled this study. We also extend our gratitude to Shanghai OE Biotech Co., Ltd. for their technical support in RNA sequencing and bioinformatic analysis.

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
4°C/-20°C/-80°C RefrigeratorHaier, ChinaHYC-390/DW-25L262/DW-86L728J
AgarHUSHI, China10000561
Agilent 2100 BioanalyzerAgilent Technologies, United StatesG2939BA
Alizarin Red S Sigma-Aldrich, United StatesA5533
Biosafety Cabinet Thermo Fisher Scientific, United States1300 A2 1.5 m
CaNO3·4H2OHUSHI, China10013960
CentrifugeThermo Fisher Scientific, United StatesX4R Pro 
CO2 IncubatorThermo Fisher Scientific, United StatesForma Steri-Cycle 3307 
Collagenase Type IVBiosharp, China BS076
Confocal MicroscopeZEISS, GermanyLSM 900
DC Pulse Power Supply Daedo Powertronics, KoreaDDP-500
DexamethasoneSigma-Aldrich, United StatesD4902
Digital MultimeterFluke, United StatesFLUKE-175
DMSOSigma-Aldrich, United StatesD2650
Electronic Balance Sartorius, Germany BSA224S-CW
FBS Gibco, United States10270-106
Flow CytometerThermo Fisher Scientific, United StatesA24858
IBMX (1-Methyl-3-isobutylxanthine)Sigma-Aldrich, United StatesI7018
Indomethacin  Sigma-Aldrich, United StatesI7378
InsulinSigma-Aldrich, United StatesI5500  
Inverted MicroscopeZEISS, GermanyAxio Observer 7 / ACR
ITS-GSigma-Aldrich, United StatesI3146
KClHUSHI, China10019318
KH2PO4HUSHI, China10019320
Linoleic Acid Sigma-Aldrich, United StatesL1376
Live Cell StationLive Cell Instrument, KoreaChamlide TC 
Low-glucose DMEMGibco, United States11885084
MgSO4·7H2O HUSHI, China10019322
Na2HPO4·12H2OHUSHI, China10019324
NaClHUSHI, China10019318
NanoDrop 2000 SpectrophotometerThermo Fisher Scientific, United StatesND-2000
Oil Red OSigma-Aldrich, United StatesO0625
Penicillin/StreptomycinGibco, United States15140122
PipettesEppendorf, Germany 4920000061
SYBR Premix EX TaqTakara, China RR420A
Toluidine Blue O     Sigma-Aldrich, United StatesT3260
TrisHUSHI, China10019326
TRIzolBeyotime, ChinaR0016
Trypsin-EDTAGibco, United States25200056
Upright MicroscopeZEISS, Germany415200-0000-000 
Vacuum GreaseDOW CORNING, United States1597418
Vitamin C   Sigma-Aldrich, United StatesA4544
Water Purification SystemThermo Fisher Scientific, United States GenPure xCAD Plus UF-TOC
β-Glycerophosphate Sigma-Aldrich, United StatesG9422

References

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  1. Qin, Y., et al. An update on adipose-derived stem cells for regenerative medicine: Where challenge meets opportunity. Adv Sci. 10 (20), e2207334(2023).
  2. Biniazan, F., Stoian, A., Haykal, S. Adipose-derived stem cells: Angioge....

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Tags

Adipose Derived Stem CellsElectrotaxisElectric Field StimulationAge Dependent MigrationStem Cell MigrationRNA SequencingSodium Channel ActivityPI3K Akt SignalingTissue RegenerationWound Healing

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