Atomic force microscopy combined with human pluripotent stem cell derived cardiomyocytes for biomechanical sensing

Pešl, Martin; Přibyl, Jan; Aćimović, Ivana; Vilotić, Aleksandra; Jelínková, Šárka; Salykin, Anton; Lacampagne, Alain; Dvořák, Petr; Méli, Albano C.; Skládal, Petr; Rotrekl, Vladimír

doi:10.1016/j.bios.2016.05.073

Cited by 53 publications

(52 citation statements)

References 45 publications

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“…In this condition, no movement of the DM1 BB could be seen by AFM measurements and even through conventional microscopy. This behavior was observed only in the DM1 samples and is consistent with a loss of synchronicity between the different contraction centers …”

Section: Resultssupporting

confidence: 79%

“…Due to its peculiarities, AFM is one of the few techniques that can directly access beating rate and intensity, from an entire BB under physiological conditions and under the action of several drugs. For instance, a recent work from Rotrekl's group have shown how AFM can monitor WT BBs under mechanical stress conditions, while at the same time recording the cellular calcium and electrophysiology response.…”

Section: Introductionmentioning

confidence: 99%

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AFM nano‐mechanical study of the beating profile of hiPSC‐derived cardiomyocytes beating bodies WT and DM1

Dinarelli¹,

Girasole²,

Spitalieri

et al. 2018

J of Molecular Recognition

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Myotonic Dystrophy type 1 (DM1) is the most common form of muscular dystrophy in adults, characterized by a variety of multisystemic features and associated with cardiac anomalies. Among cardiac phenomena, conduction defects, ventricular arrhythmias, and dilated cardiomyopathy represent the main cause of sudden death in DM1 patients. Patient-specific induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a powerful in vitro model for molecular, biochemical, and physiological studies of disease in the target cells. Here, we used an Atomic Force Microscope (AFM) to measure the beating profiles of a large number of cells, organized in CM clusters (Beating Bodies, BBs), obtained from wild type (WT) and DM1 patients. We monitored the evolution over time of the frequency and intensity of the beating. We determined the variations between different BBs and over various areas of a single BB, caused by morphological and biomechanical variations. We exploited the AFM tip to apply a controlled force over the BBs, to carefully assess the biomechanical reaction of the different cell clusters over time, both in terms of beating frequency and intensity. Our measurements demonstrated differences between the WT and DM1 clusters highlighting, for the DM1 samples, an instability which was not observed in WT cells. We measured differences in the cellular response to the applied mechanical stimulus in terms of beating synchronicity over time and cell tenacity, which are in good agreement with the cellular behavior in vivo. Overall, the combination of hiPSC-CMs with AFM characterization can become a new tool to study the collective movements of cell clusters in different conditions and can be extended to the characterization of the BB response to chemical and pharmacological stimuli.

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Section: Resultssupporting

confidence: 79%

Section: Introductionmentioning

confidence: 99%

AFM nano‐mechanical study of the beating profile of hiPSC‐derived cardiomyocytes beating bodies WT and DM1

Dinarelli¹,

Girasole²,

Spitalieri

et al. 2018

J of Molecular Recognition

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show abstract

“…Despite the complications mentioned in the previous chapter, our group has successfully developed reliable uniformly sized EB models, containing hPSC‐CMs . These cellular constructs of cardiac syncytium were coupled to an atomic force microscopy (AFM) force sensor to perform a high‐fidelity contraction pattern as an hPSC‐CM–based biosensor . The high consistency in the cluster of cells and its constant size among different cell lines allow a comparable and stable beating pattern.…”

Section: Methods Results and Discussionmentioning

confidence: 99%

“…The ability of AFM to evaluate the bending of the cantilever probe with extremely high precision allows this device to be used as a mechanical nanosensor or as a micromechanical transducer for the construction of complex nanobiosensors, such as in the FluidFM modification by Ossola et al Atomic force microscopy as a base for construction of a biosensor device, when combined with bio‐objects as a recognition part, was used as a mechanical sensor for cell penetration by nanoneedles, in cell‐based biosensing of drug effects and in the study of grow factor effects . The use of AFM to study mechanobiological properties was demonstrated in single cells and clusters . Lieu and coworkers have used AFM in synergy with Ca 2 + imaging to prove that hESC‐CMs lack T‐tubule structures, fundamental in adult CMs for the proper Ca 2 + ‐induced Ca 2 + release mechanism .…”

Section: Methods Results and Discussionmentioning

confidence: 99%

“…16 These cellular constructs of cardiac syncytium were coupled to an atomic force microscopy (AFM) force sensor to perform a high-fidelity contraction pattern as an hPSC-CM-based biosensor. 74 The high consistency in the cluster of cells and its constant size among different cell lines allow a comparable and stable beating pattern. This system allows for force-and rhythm-related drug effect tracking, but also disease model application and drug study in specific inherited diseases, hampering force and rhythm.…”

Section: Methods Results and Discussionmentioning

confidence: 99%

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Phenotypic assays for analyses of pluripotent stem cell–derived cardiomyocytes

Pešl

Přibyl

Caluori

et al. 2016

J of Molecular Recognition

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Stem cell-derived cardiomyocytes (CMs) hold great hopes for myocardium regeneration because of their ability to produce functional cardiac cells in large quantities. They also hold promise in dissecting the molecular principles involved in heart diseases and also in drug development, owing to their ability to model the diseases using patient-specific human pluripotent stem cell (hPSC)-derived CMs. The CM properties essential for the desired applications are frequently evaluated through morphologic and genotypic screenings. Even though these characterizations are necessary, they cannot in principle guarantee the CM functionality and their drug response. The CM functional characteristics can be quantified by phenotype assays, including electrophysiological, optical, and/or mechanical approaches implemented in the past decades, especially when used to investigate responses of the CMs to known stimuli (eg, adrenergic stimulation). Such methods can be used to indirectly determine the electrochemomechanics of the cardiac excitation-contraction coupling, which determines important functional properties of the hPSC-derived CMs, such as their differentiation efficacy, their maturation level, and their functionality. In this work, we aim to systematically review the techniques and methodologies implemented in the phenotype characterization of hPSC-derived CMs. Further, we introduce a novel approach combining atomic force microscopy, fluorescent microscopy, and external electrophysiology through microelectrode arrays. We demonstrate that this novel method can be used to gain unique information on the complex excitation-contraction coupling dynamics of the hPSC-derived CMs.

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Advances in Biosensors and Diagnostic Technologies Using Nanostructures and Nanomaterials

Welch

Powell

Clevinger

et al. 2021

Adv Funct Materials

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Nanoscale materials have unique properties that make them especially useful for biomedical diagnostic applications. Recent developments in nanoengineering have resulted in increasing use of nanostructures in biosensors. Various types of 0D, 1D, 2D, and 3D nanostructures have been used to improve biosensor sensitivity, selectivity, limit of detection, and time to result, among other metrics. These nanostructures have been integrated into electrochemical, optical, and other biosensors for this purpose. Here, the most recent advances in the use of nanostructured materials in biosensors are described. This includes a discussion of nanoparticles, nanorods, nanofibers, nanopillars, nanowires, nanosheets, indented nanopatterns (nanoholes and nanoslits), nanogaps, nanochannels, nanopores, nanofunctionalized surfaces, and complex hierarchical structures and their unique advantages and applications in biosensors. Clinical applications of these nanobiosensors are also highlighted along with a discussion of the future directions for these materials in diagnostics.

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Atomic force microscopy combined with human pluripotent stem cell derived cardiomyocytes for biomechanical sensing

Cited by 53 publications

References 45 publications

AFM nano‐mechanical study of the beating profile of hiPSC‐derived cardiomyocytes beating bodies WT and DM1

AFM nano‐mechanical study of the beating profile of hiPSC‐derived cardiomyocytes beating bodies WT and DM1

Phenotypic assays for analyses of pluripotent stem cell–derived cardiomyocytes

Advances in Biosensors and Diagnostic Technologies Using Nanostructures and Nanomaterials

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