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* These authors contributed equally
Here, we present a protocol for applying nanosecond pulse electric field (nsPEF) to stimulate Schwann cells in vitro. The synthesis and secretion ability of relevant factors and cell behavior changes validated the successful stimulation using nsPEF. The study gives a positive view of the peripheral nerve regeneration method.
Schwann cells (SCs) are myelinating cells of the peripheral nervous system, playing a crucial role in peripheral nerve regeneration. Nanosecond Pulse Electric Field (nsPEF) is an emerging method applicable in nerve electrical stimulation that has been demonstrated to be effective in stimulating cell proliferation and other biological processes. Aiming to assess whether SCs undergo significant changes under nsPEF and help explore the potential for new peripheral nerve regeneration methods, cultured RSC96 cells were subjected to nsPEF stimulation at 5 kV and 10 kV, followed by continued cultivation for 3-4 days. Subsequently, some relevant factors expressed by SCs were assessed to demonstrate the successful stimulation, including the specific marker protein, neurotrophic factor, transcription factor, and myelination regulator. The representative results showed that nsPEF significantly enhanced the proliferation and migration of SCs and the ability to synthesize relevant factors that contribute positively to the regeneration of peripheral nerves. Simultaneously, lower expression of GFAP indicated the benign prognosis of peripheral nerve injuries. All these outcomes show that nsPEF has great potential as an efficient treatment method for peripheral nerve injuries by stimulating SCs.
Each year, millions of people are affected by nerve injuries involving both the peripheral nervous system (PNS) and the central nervous system (CNS)1. Studies have demonstrated that the axonal repair capacity of the CNS is quite limited after nerve injuries, while the PNS shows enhanced capacity due to the significant plasticity of SCs2. Nevertheless, achieving complete regeneration after peripheral nerve injuries remains arduous and continues to pose a significant challenge to human health3,4. Nowadays, autografts have remained a common treatment despite the dra....
1. Thawing of cryopreserved RSC96 cells
Low-intensity pulsed electric fields stimulate cell proliferation
According to the CCK-8 assay, the proliferation rate of RSC96 in the 5 kV/cm group was significantly faster than that of the control group cells. However, as the parameters increased (20 kV/cm and 40 kV/cm), the proliferation rate was unstable, even lower than that of the control group. The cell proliferation rate of RSC96 cells in the 40 kV/cm group was significantly lower than the control and 5 kV/cm groups, showing a significant s.......
In recent years, the application of nsPEF has experienced boosting growth, as reported. nsPEF has a highly targeted effect on only the desired area, providing enough energy to treat without causing additional thermal damage, making it safer for the human body28. These characteristics give it promising translational prospects in tumor treatment and nerve regeneration. However, some studies have proposed some limitations of nsPEF. Compared with materials research, ES is constrained by external power.......
This work was funded by the National Key Scientific Instrument and Equipment Development Project (NO.82027803).
....Name | Company | Catalog Number | Comments |
Antifade mounting medium | Wuhan Xavier Biotechnology Co., LTD | G1401 | |
Anti-GFAP Mouse mAb | Wuhan Xavier Biotechnology Co., LTD | GB12100-100 | |
Anti-Neurofilament heavy polypeptide Mouse mAb | Wuhan Xavier Biotechnology Co., LTD | GB12144-100 | |
Anti-S100 beta Mouse mAb | Wuhan Xavier Biotechnology Co., LTD | GB14146-100 | |
BSA | Wuhan Xavier Biotechnology Co., LTD | GC305010 | |
Coverslip | Jiangsu Shitai experimental equipment Co., LTD | 10212432C | |
CY3-labeled goat anti-mouse IgG | Wuhan Xavier Biotechnology Co., LTD | GB21302 | |
DAPI Staining Reagent | Wuhan Xavier Biotechnology Co., LTD | G1012 | |
Decolorizing shaker | Wuhan Xavier Biotechnology Co., LTD | DS-2S100 | |
High Voltage Power Supply for nsPEF | Matsusada Precision Inc. | AU-60P1.6-L | |
Histochemical pen | Wuhan Xavier Biotechnology Co., LTD | G6100 | |
Membrane breaking liquid | Wuhan Xavier Biotechnology Co., LTD | G1204 | |
Microscope slide | Wuhan Xavier Biotechnology Co., LTD | G6012 | |
Palm centrifuge | Wuhan Xavier Biotechnology Co., LTD | MS6000 | |
PBS powdered | Wuhan Xavier Biotechnology Co., LTD | G0002 | |
Pipette | Wuhan Xavier Biotechnology Co., LTD | ||
Positive fluorescence microscope | Nikon, Japan | NIKON ECLIPSE C1 | |
Rabbit Anti-SOX10/AF488 Conjugated antibody | Beijing Bioss Biotechnology Co., LTD | BS-20563R-AF488 | |
RSC96 Schwann cells | Wuhan Xavier Biotechnology Co., LTD | STCC30007G-1 | |
scanister | 3DHISTECH | Pannoramic MIDI | |
Special cable for nsPEF | Times Microwave Systems | M17/78-RG217 | |
Turbine mixer | Wuhan Xavier Biotechnology Co., LTD | MV-100 |
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