On-chip electroporation, membrane repair dynamics and transient in-cell recordings by arrays of gold mushroom-shaped microelectrodes

Hai, Aviad; Spira, Micha Ε.

doi:10.1039/c2lc40091j

Cited by 84 publications

(96 citation statements)

References 51 publications

(68 reference statements)

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“…Thereafter, cell biological repair processes sealed off the electroporated nanopores50 and the intracellularly recorded action potential gradually reverted to the typical shaped extracellular FP (Fig. 6)3451.…”

Section: Resultsmentioning

confidence: 97%

On-chip, multisite extracellular and intracellular recordings from primary cultured skeletal myotubes

Rabieh

Ojovan

Shmoel

et al. 2016

Sci Rep

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In contrast to the extensive use of microelectrode array (MEA) technology in electrophysiological studies of cultured neurons and cardiac muscles, the vast field of skeletal muscle research has yet to adopt the technology. Here we demonstrate an empowering MEA technology for high quality, multisite, long-term electrophysiological recordings from cultured skeletal myotubes. Individual rat skeletal myotubes cultured on micrometer sized gold mushroom-shaped microelectrode (gMμE) based MEA tightly engulf the gMμEs, forming a high seal resistance between the myotubes and the gMμEs. As a consequence, spontaneous action potentials generated by the contracting myotubes are recorded as extracellular field potentials with amplitudes of up to 10 mV for over 14 days. Application of a 10 ms, 0.5–0.9 V voltage pulse through the gMμEs electroporated the myotube membrane, and transiently converted the extracellular to intracellular recording mode for 10–30 min. In a fraction of the cultures stable attenuated intracellular recordings were spontaneously produced. In these cases or after electroporation, subthreshold spontaneous potentials were also recorded. The introduction of the gMμE-MEA as a simple-to-use, high-quality electrophysiological tool together with the progress made in the use of cultured human myotubes opens up new venues for basic and clinical skeletal muscle research, preclinical drug screening, and personalized medicine.

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Section: Resultsmentioning

confidence: 97%

On-chip, multisite extracellular and intracellular recordings from primary cultured skeletal myotubes

Rabieh

Ojovan

Shmoel

et al. 2016

Sci Rep

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“…6D), enough time to allow non-critical sized cell membrane pores to heal25. Measurements taken at 6 hours (Fig.…”

Section: Resultsmentioning

confidence: 99%

Biomimetic surface patterning for long-term transmembrane access

VanDersarl

Renaud

2016

Sci Rep

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Here we present a planar patch clamp chip based on biomimetic cell membrane fusion. This architecture uses nanometer length-scale surface patterning to replicate the structure and function of membrane proteins, creating a gigaohm seal between the cell and a planar electrode array. The seal is generated passively during cell spreading, without the application of a vacuum to the cell surface. This interface can enable cell-attached and whole-cell recordings that are stable to 72 hours, and generates no visible damage to the cell. The electrodes can be very small (<5 μm) and closely packed, offering a high density platform for cellular measurement.

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“…The nanopillar electrode array platform recently developed by our group and others [15][16][17] offers the opportunity to measure, in multiplex, intracellular action potentials of single cardiomyocytes in a beating, confluent monolayer. This measurement could yield the information relevant to pre-clinically monitor drug cardiotoxicity 3 , track cardiomyocyte maturation, and perform any variety of experiments enabled by multi-day current clamp-like measurements.…”

Section: Introductionmentioning

confidence: 99%

Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

et al. 2017

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The measurement of the electrophysiology of human pluripotent stem cell-derived cardiomyocytes is critical for their biomedical applications, from disease modeling to drug screening. Yet, a method that enables the high-throughput intracellular electrophysiology measurement of single cardiomyocytes in adherent culture is not available. To address this area, we have fabricated vertical nanopillar electrodes that can record intracellular action potentials from up to 60 single beating cardiomyocytes. Intracellular access is achieved by highly localized electroporation, which allows for low impedance electrical access to the intracellular voltage. Herein, we demonstrate that this method provides the accurate measurement of the shape and duration of intracellular action potentials, validated by patch clamp, and can facilitate cellular drug screening and disease modeling using human pluripotent stem cells. This study validates the use of nanopillar electrodes for myriad further applications of human pluripotent stem cell-derived cardiomyocytes such as cardiomyocyte maturation monitoring and electrophysiology-contractile force correlation. There have been many efforts to generate cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) due to the difficulty in obtaining human cardiac tissue by other methods 7 . In the last decade, efficient and reliable protocols, such as the embryoid body method and the monolayer differentiation method have been established to generate hPSC-CMs within 2 weeks and with 470% efficiency 8,9 . Critically, hPSC-CMs exhibit key cardiomyocyte properties, expressing the expected ion channels, exhibiting human cardiac-type action potentials, and containing functional sarcomeres.Despite their potential, applications of hPSC-CMs have been hampered by the cells' intrinsic heterogeneity and the quality of functional assays for monitoring their electrophysiology. First, hPSC-CMs are intrinsically heterogeneous, consisting of three subpopulations: atrial-, ventricular-, and nodal-like 10 . Furthermore, electrophysiology measurements have shown that the shapes of action potentials vary significantly between studies and within studies among different cell lines and differentiation methods 11,12 . Therefore, the high heterogeneity among hPSC-CMs requires that a large number of cells be analyzed to generate statistically meaningful conclusions.The gold standard for measuring cardiomyocyte electrophysiology is manual patch clamp; however, this technique is laborious

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On-chip electroporation, membrane repair dynamics and transient in-cell recordings by arrays of gold mushroom-shaped microelectrodes

Cited by 84 publications

References 51 publications

On-chip, multisite extracellular and intracellular recordings from primary cultured skeletal myotubes

On-chip, multisite extracellular and intracellular recordings from primary cultured skeletal myotubes

Biomimetic surface patterning for long-term transmembrane access

Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

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