The MyoPulser field stimulator, a do it yourself programmable electronic pacemaker for contracting cells and tissues

Ott, Christiane; Jung, Tobias

doi:10.1038/s41598-023-29145-3

Cited by 4 publications

(1 citation statement)

References 27 publications

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“…Furthermore, our computational models confirm that the TMV response is influenced by electoporation already at the lowest electric fields used in experiments, which were close to the stimulation threshold. Conventionally, in vitro electrostimulation is done with millisecond pulses 63 . Our results suggest that when applying 100 µs pulses, as used conventionally in electroporation applications, electroporation may exist already at electric fields close to the stimulation thresholds.…”

Section: Discussionmentioning

confidence: 99%

Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells

Batista Napotnik,

Kos,

Jarm

et al. 2024

Sci Rep

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Electric pulses used in electroporation-based treatments have been shown to affect the excitability of muscle and neuronal cells. However, understanding the interplay between electroporation and electrophysiological response of excitable cells is complex, since both ion channel gating and electroporation depend on dynamic changes in the transmembrane voltage (TMV). In this study, a genetically engineered human embryonic kidney cells expressing NaV1.5 and Kir2.1, a minimal complementary channels required for excitability (named S-HEK), was characterized as a simple cell model used for studying the effects of electroporation in excitable cells. S-HEK cells and their non-excitable counterparts (NS-HEK) were exposed to 100 µs pulses of increasing electric field strength. Changes in TMV, plasma membrane permeability, and intracellular Ca2+ were monitored with fluorescence microscopy. We found that a very mild electroporation, undetectable with the classical propidium assay but associated with a transient increase in intracellular Ca2+, can already have a profound effect on excitability close to the electrostimulation threshold, as corroborated by multiscale computational modelling. These results are of great relevance for understanding the effects of pulse delivery on cell excitability observed in context of the rapidly developing cardiac pulsed field ablation as well as other electroporation-based treatments in excitable tissues.

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Section: Discussionmentioning

confidence: 99%

Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells

Batista Napotnik,

Kos,

Jarm

et al. 2024

Sci Rep

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Optimization of H9c2 differentiation leads to calcium-active and striated cardiac cells without addition of retinoic acid

Brock,

Hörning

2024

Preprint

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As a reliable alternative to animal testing in cardiovascular research, it is crucial to improve differentiation of immortalized cell lines. In this study, we focused on optimizing the differentiation efficiency of the H9c2 cell line into cardiomyocytes using a high-throughput, automated image processing approach. While previous studies used protocols involving retinoic acid to enhance cardiac differentiation, we applied a simplified medium composition that results in higher differentiation rates. Along that line, we differentiated H9c2 cells into cardiomyocytes, which not only showed sarcomerecharacteristic striation but also periodic intracellular calcium signaling for the first time. As a second step, we examined the potential application of polyacrylamide hydrogels (E = 12 kPa) with de- fined fibronectin coating densities. The optimum fibronectin density of 2.6 μg/cm2 found for single cells was investigated to further improve the dif- ferentiation efficiency. However, the differentiation and proliferation dynamics dominate the adhesion forces between the cells and the hydrogel, and thus, result in premature clustering and detachment. In conclusion, we identified an optimized differentiation protocol and provided a basis for the further investigation necessary to potentially use hydrogels as natural cell environment, aiming to raise the differentiation efficiency even more.

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Optimization of H9c2 differentiation leads to calcium-active and striated cardiac cells without addition of retinoic acid

Brock,

Hörning

2024

Front. Cell Dev. Biol.

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As a reliable alternative to animal testing in cardiovascular research, it is crucial to improve differentiation of immortalized cell lines. In this study, we focused on optimizing the differentiation efficiency of the H9c2 cell line into cardiomyocytes using a high-throughput, automated image processing approach. While previous studies used protocols involving retinoic acid to enhance cardiac differentiation, we applied a simplified medium composition that results in higher differentiation rates. Along that line, we differentiated H9c2 cells into cardiomyocytes, which not only showed sarcomere-characteristic striation but also periodic intracellular calcium signaling for the first time. As a second step, we examined the potential application of polyacrylamide hydrogels (E=12 kPa) with defined fibronectin coating densities. The optimum fibronectin density of 2.6 μg/cm2 found for single cells was investigated to further improve the differentiation efficiency. However, the differentiation and proliferation dynamics dominate the adhesion forces between the cells and the hydrogel, and thus, result in premature clustering and detachment. In conclusion, we identified an optimized differentiation protocol and provided a basis for the further investigation necessary to potentially use hydrogels as natural cell environment, aiming to raise the differentiation efficiency even more.

show abstract

The MyoPulser field stimulator, a do it yourself programmable electronic pacemaker for contracting cells and tissues

Cited by 4 publications

References 27 publications

Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells

Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells

Optimization of H9c2 differentiation leads to calcium-active and striated cardiac cells without addition of retinoic acid

Optimization of H9c2 differentiation leads to calcium-active and striated cardiac cells without addition of retinoic acid

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