Constant-potential environment for activating and synchronizing cardiomyocyte colonies with on-chip ion-depleting perm-selective membranes

Yadav, Vivek; Chong, Nicholas; Ellis, Bradley W.; Ren, Xiang; Senapati, Satyajyoti; Zorlutuna, Pınar

doi:10.1039/d0lc00809e

Cited by 7 publications

(4 citation statements)

References 74 publications

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“…48 Low-frequency AC electric fields can also influence the cell membrane action potential and open VGCCs, allowing for an influx of Ca 2+ ions into the cell. 57 Alongside piezoelectric stimulation, audible acoustic stimulation can activate mechanosensing calcium channels such as Piezo-1 through mechanical force induction. 58,59 Overall, we found that increased metabolite levels coupled with enhanced intracellular calcium levels potentially activated multiple stages of sEV production in both cell lines.…”

Section: Enhanced Sev Production In Stimulated Pess Via Increased Cel...mentioning

confidence: 99%

Stimulative piezoelectric nanofibrous scaffolds for enhanced small extracellular vesicle production in 3D cultures

Johnston,

Jeon,

Choi

et al. 2024

Preprint

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Small extracellular vesicles (sEVs) have great promise as effective carriers for drug delivery. However, the challenges associated with the efficient production of sEVs hinder their clinical applications. Herein, we report a stimulative 3D culture platform for enhanced sEV production. The proposed platform consists of a piezoelectric nanofibrous scaffold (PES) coupled with acoustic stimulation to enhance sEV production of cells in a 3D biomimetic microenvironment. Combining cell stimulation with a 3D culture platform in this stimulative PES enables a 49 fold increase in the production rate per cell with minimal deviations in particle size and protein composition compared with standard 2D cultures. We find that the enhanced sEV production is attributable to the activation and upregulation of crucial sEV production steps through the synergistic effect of stimulation and the 3D microenvironment. Moreover, changes in cell morphology lead to cytoskeleton redistribution through cell matrix interactions in the 3D cultures. This in turn facilitates intracellular EV trafficking, which impacts the production rate. Overall, our work provides a promising 3D cell culture platform based on piezoelectric biomaterials for enhanced sEV production. This platform is expected to accelerate the potential use of sEVs for drug delivery and broad biomedical applications.

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Section: Enhanced Sev Production In Stimulated Pess Via Increased Cel...mentioning

confidence: 99%

Stimulative piezoelectric nanofibrous scaffolds for enhanced small extracellular vesicle production in 3D cultures

Johnston,

Jeon,

Choi

et al. 2024

Preprint

Self Cite

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show abstract

“…A mixture of the curing agent and polydimethylsiloxane (PDMS, Sylgard 184, Dow Corning) base, with a weight ratio of 1:10, was prepared and degassed for one hour to eliminate air bubbles (Yadav et al, 2020). Subsequently, the PDMS was spin-coated onto a 50 x 75 mm glass slide and left overnight for curing, forming a thin film of approximately 200µm.…”

Section: Device Fabricationmentioning

confidence: 99%

A Mem-dELISA platform for dual color and ultrasensitive digital detection of colocalized proteins on extracellular vesicles

Sharma,

Yadav,

Burchett

et al. 2024

Preprint

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Accurate, multiplex, and ultrasensitive measurement of different colocalized protein markers on individual tumor-derived extracellular vesicles (EVs) and dimerized proteins with multiple epitopes could provide insights into cancer heterogeneity, therapy management and early diagnostics that cannot be extracted from bulk methods. However, current digital protein assays lack certain features to enable robust colocalization, including multi-color detection capability, large dynamic range, and selectivity against background proteins. Here, we report a lithography-free, inexpensive (< $0.1) and ultrasensitive dual-color Membrane Digital ELISA (Mem-dELISA) platform by using track-etched polycarbonate (PCTE) membranes to overcome these shortcomings. Their through-pores remove air bubbles through wicking before they are sealed on one side by adhesion to form microwells. Immunomagnetic bead-analyte complexes and substrate solution are then loaded into the microwells from the opposite side, with >80% loading efficiency, before sealing with oil. This enables duplex digital protein colorimetric assay with beta galactosidase and alkaline phosphatase enzymes. The platform achieves 5 logs of dynamic range with a limit of detection of 10 aM for both Biotinylated β-galactosidase (B-βG) and Biotin Alkaline Phosphatase Conjugated (B-ALP) proteins. We demonstrate its potential by showing that a higher dosage of paclitaxel suppresses EpCAM-positive EVs but not GPC-1 positive EVs from breast cancer cells, a decline in chemo-resistance that cannot be detected with Western blot analysis of cell lysate. The Mem-dELISA is poised to empower researchers to conduct ultrasensitive, high throughput protein colocalization studies for disease diagnostics, treatment monitoring and biomarker discovery.

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“…Concretely, CMs can be obtained from animal or human hearts or by the differentiation process from stem cells. In general terms, CMs from primary culture have some limitations for the study of cardiac physiology, such as low proliferative capacity and the absence of spontaneous beat, among others [ 19 , 20 , 21 , 22 , 23 ]. As an alternative, differentiated CMs derived from pluripotent cells, mesenchymal stem cells (MSCs), embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs), are essential for the development of MI models [ 24 , 25 , 26 , 27 , 28 ].…”

Section: Experimental Models Of Cardiac Regenerationmentioning

confidence: 99%

Comparative Analysis of Heart Regeneration: Searching for the Key to Heal the Heart—Part I: Experimental Injury Models to Study Cardiac Regeneration

Castillo-Casas,

Caño-Carrillo,

Sánchez-Fernández

et al. 2023

JCDD

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Cardiovascular diseases are the leading cause of death worldwide, among which, ischemic heart disease is the most prevalent. Myocardial infarction results from occlusion of a coronary artery, which leads to an insufficient blood supply to the myocardium. As is well known, the massive loss of cardiomyocytes cannot be solved due the limited regenerative ability of the adult mammalian heart. In contrast, some lower vertebrate species can regenerate the heart after injury; their study has disclosed some of the involved cell types, molecular mechanisms and signaling pathways during the regenerative process. In this two-part review, we discuss the current state of the principal response in heart regeneration, where several involved processes are essential for full cardiac function in recovery.

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Constant-potential environment for activating and synchronizing cardiomyocyte colonies with on-chip ion-depleting perm-selective membranes

Abstract: In this study, an ion depleted zone created by an ion-selective membrane was used to impose a high and uniform constant extracellular potential over an entire ~1000-cell rat cardiomyocyte (rCM)...

Cited by 7 publications

References 74 publications

Stimulative piezoelectric nanofibrous scaffolds for enhanced small extracellular vesicle production in 3D cultures

Stimulative piezoelectric nanofibrous scaffolds for enhanced small extracellular vesicle production in 3D cultures

A Mem-dELISA platform for dual color and ultrasensitive digital detection of colocalized proteins on extracellular vesicles

Comparative Analysis of Heart Regeneration: Searching for the Key to Heal the Heart—Part I: Experimental Injury Models to Study Cardiac Regeneration

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