Electric pulses: a flexible tool to manipulate cytosolic calcium concentrations and generate spontaneous-like calcium oscillations in mesenchymal stem cells

Ménorval, Marie-Amélie de; André, Franck; Silve, Aude; Dalmay, Claire; Français, Olivier; Pioufle, Bruno Le; Mir, Lluis M.

doi:10.1038/srep32331

Cited by 22 publications

(35 citation statements)

References 40 publications

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“…The ultrashort pulses (one thousandth to one ten thousandth shorter than the pulses of 100 μs used in our study) called nanopulses or nanosecond pulsed electric fields (nsPEFs) have been used already to induce Ca 2+ bursts in the cell, which resulted from the electropermeabilization of the ER, the plasma membrane or both [13, 30]. However, contrary to the 100-μs electric pulses, the technology to deliver nanopulses is not yet spread in the laboratories and is not simple to use.…”

Section: Discussionmentioning

confidence: 99%

Electrical control of calcium oscillations in mesenchymal stem cells using microsecond pulsed electric fields

2017

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BackgroundHuman mesenchymal stem cells are promising tools for regenerative medicine due to their ability to differentiate into many cellular types such as osteocytes, chondrocytes and adipocytes amongst many other cell types. These cells present spontaneous calcium oscillations implicating calcium channels and pumps of the plasma membrane and the endoplasmic reticulum. These oscillations regulate many basic functions in the cell such as proliferation and differentiation. Therefore, the possibility to mimic or regulate these oscillations might be useful to regulate mesenchymal stem cells biological functions.MethodsOne or several electric pulses of 100 μs were used to induce Ca2+ spikes caused by the penetration of Ca2+ from the extracellular medium, through the transiently electropermeabilized plasma membrane, in human adipose mesenchymal stem cells from several donors. Attached cells were preloaded with Fluo-4 AM and exposed to the electric pulse(s) under the fluorescence microscope. Viability was also checked.ResultsAccording to the pulse(s) electric field amplitude, it is possible to generate a supplementary calcium spike with properties close to those of calcium spontaneous oscillations, or, on the contrary, to inhibit the spontaneous calcium oscillations for a very long time compared to the pulse duration. Through that inhibition of the oscillations, Ca2+ oscillations of desired amplitude and frequency could then be imposed on the cells using subsequent electric pulses. None of the pulses used here, even those with the highest amplitude, caused a loss of cell viability.ConclusionsAn easy way to control Ca2+ oscillations in mesenchymal stem cells, through their cancellation or the addition of supplementary Ca2+ spikes, is reported here. Indeed, the direct link between the microsecond electric pulse(s) delivery and the occurrence/cancellation of cytosolic Ca2+ spikes allowed us to mimic and regulate the Ca2+ oscillations in these cells. Since microsecond electric pulse delivery constitutes a simple technology available in many laboratories, this new tool might be useful to further investigate the role of Ca2+ in human mesenchymal stem cells biological processes such as proliferation and differentiation.

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Section: Discussionmentioning

confidence: 99%

Electrical control of calcium oscillations in mesenchymal stem cells using microsecond pulsed electric fields

2017

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“…In bovine chromaffin cells, a single 5-ns, 50 kV/cm PEF caused Ca 2+ mobilization which depended on L-type VGCC and was mediated by the tetrodotoxin-insensitive Na + uptake; the data argued against direct activation of VGSC by nsPEF 26 . Other studies with 10-ns pulses did not even consider possible activation of VGSC and regarded Ca 2+ mobilization solely as a result of electroporation 27 .…”

Section: Introductionmentioning

confidence: 99%

Damage-free peripheral nerve stimulation by 12-ns pulsed electric field

Casciola

Xiao

Pakhomov

2017

Sci Rep

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Modern technologies enable deep tissue focusing of nanosecond pulsed electric field (nsPEF) for non-invasive nerve and muscle stimulation. However, it is not known if PEF orders of magnitude shorter than the activation time of voltage-gated sodium channels (VGSC) would evoke action potentials (APs). One plausible scenario requires the loss of membrane integrity (electroporation) and resulting depolarization as an intermediate step. We report, for the first time, that the excitation of a peripheral nerve can be accomplished by 12-ns PEF without electroporation. 12-ns stimuli at 4.1–11 kV (3.3–8.8 kV/cm) evoked APs similarly to conventional stimuli (100–250 μs, 1–5 V, 103–515 V/m), except for having higher selectivity for the faster nerve fibers. Nerves sustained repeated tetanic stimulations (50 Hz or 100 Hz for 1 min) alternately by 12-ns PEF and by conventional pulses. Such tetani caused a modest AP decrease, to a similar extent for both types of stimuli. Nerve refractory properties were not affected. The lack of cumulative damages even from tens of thousands of 12-ns stimuli and the similarities with the conventional stimulation prove VGSC activation by nsPEF without nerve membrane damage.

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“…The application to cells of one or several nsPEF of 10 ns duration is very common in nanoporation [10–15]. Different types of applicators and devices can be used without compromising the shape of the applied pulse [16–21].…”

Section: Introductionmentioning

confidence: 99%

Technological and Theoretical Aspects for Testing Electroporation on Liposomes

Denzi

Valle

Esposito

et al. 2017

BioMed Research International

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Recently, the use of nanometer liposomes as nanocarriers in drug delivery systems mediated by nanoelectroporation has been proposed. This technique takes advantage of the possibility of simultaneously electroporating liposomes and cell membrane with 10-nanosecond pulsed electric fields (nsPEF) facilitating the release of the drug from the liposomes and at the same time its uptake by the cells. In this paper the design and characterization of a 10 nsPEF exposure system is presented, for liposomes electroporation purposes. The design and the characterization of the applicator have been carried out choosing an electroporation cuvette with 1 mm gap between the electrodes. The structure efficiency has been evaluated at different experimental conditions by changing the solution conductivity from 0.25 to 1.6 S/m. With the aim to analyze the influence of device performances on the liposomes electroporation, microdosimetric simulations have been performed considering liposomes of 200 and 400 nm of dimension with different inner and outer conductivity (from 0.05 to 1.6 S/m) in order to identify the voltage needed for their poration.

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Electric pulses: a flexible tool to manipulate cytosolic calcium concentrations and generate spontaneous-like calcium oscillations in mesenchymal stem cells

Cited by 22 publications

References 40 publications

Electrical control of calcium oscillations in mesenchymal stem cells using microsecond pulsed electric fields

Electrical control of calcium oscillations in mesenchymal stem cells using microsecond pulsed electric fields

Damage-free peripheral nerve stimulation by 12-ns pulsed electric field

Technological and Theoretical Aspects for Testing Electroporation on Liposomes

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