A Pt layer/Pt disk electrode configuration to evaluate respiration and alkaline phosphatase activities of mouse embryoid bodies

Obregón, Raquel; Horiguchi, Yoshiko; Arai, Toshiharu; Abe, Shihomi; Zhou, Yifei; RyosukeTakahashi,; Hisada, Akiko; Ino, Kosuke; Shiku, Hitoshi; Matsue, Tomokazu

doi:10.1016/j.talanta.2012.01.059

Cited by 24 publications

(26 citation statements)

References 32 publications

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“…Interestingly, SECM can detect more rapidly changes in cellular activity than conventional live/dead stains [9]. Besides, intracellular measurements of the cell redox activity [10], [11] were reported as well as assays to evaluate the differentiation status of embryonic stem cells using an indirect enzymatic assay [12], [13].…”

Section: Introductionmentioning

confidence: 99%

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

et al. 2014

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While scanning electrochemical microscopy (SECM) is a powerful technique for non-invasive analysis of cells, SECM-based assays remain scarce and have been mainly limited so far to single cells, which is mostly due to the absence of suitable platform for experimentation on 3D cellular aggregates or microtissues. Here, we report stamping of a Petri dish with a microwell array for large-scale production of microtissues followed by their in situ analysis using SECM. The platform is realized by hot embossing arrays of microwells (200 μm depth; 400 μm diameter) in commercially available Petri dishes, using a PDMS stamp. Microtissues form spontaneously in the microwells, which is demonstrated here using various cell lines (e.g., HeLa, C2C12, HepG2 and MCF-7). Next, the respiratory activity of live HeLa microtissues is assessed by monitoring the oxygen reduction current in constant height mode and at various distances above the platform surface. Typically, at a 40 μm distance from the microtissue, a 30% decrease in the oxygen reduction current is measured, while above 250 μm, no influence of the presence of the microtissues is detected. After exposure to a model drug (50% ethanol), no such changes in oxygen concentration are found at any height in solution, which reflects that microtissues are not viable anymore. This is furthermore confirmed using conventional live/dead fluorescent stains. This live/dead assay demonstrates the capability of the proposed approach combining SECM and microtissue arrays formed in a stamped Petri dish for conducting cellular assays in a non-invasive way on 3D cellular models.

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Section: Introductionmentioning

confidence: 99%

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

et al. 2014

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“…3B). 14 To incorporate these electrodes into a single probe device, a Pt pseudo-reference was prepared on the surface of the probe device. The device was successfully applied to the evaluation of O 2 consumption of a single embryoid body (EB) from mouse embryonic stem (ES) cells.…”

Section: Evaluation Of O 2 Consumption Of 3d Cultured Cellsmentioning

confidence: 99%

“…5A). 14,25,26 Additionally, SECM was applied in the non-invasive monitoring of osteogenic differentiation of C2C12 microtissues. 27 Differentiation was assessed by measuring the activity of ALP, which is known to be highly expressed during differentiation.…”

Section: Evaluation Of Endogenous Enzyme Activity Of 3d Cultured Cellsmentioning

confidence: 99%

Micro/nanoelectrochemical probe and chip devices for evaluation of three-dimensional cultured cells

Ino

Şen

Shiku

et al. 2017

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Herein, we present an overview of recent research progress in the development of micro/nanoelectrochemical probe and chip devices for the evaluation of three-dimensional (3D) cultured cells. First, we discuss probe devices: a general outline, evaluation of O 2 consumption, enzyme-modified electrodes, evaluation of endogenous enzyme activity, and the collection of cell components from cell aggregates are discussed. The next section is focused on integrated chip devices: a general outline, electrode array devices, smart electrode array devices, droplet detection of 3D cultured cells, cell manipulation using dielectrophoresis (DEP), and electrodeposited hydrogels used for fabrication of 3D cultured cells on chip devices are discussed. Finally, we provide a summary and discussion of future directions of research in this field.

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“…We have previously reported that electrochemical methods can be applied for the evaluation of ES cell differentiation via their ALP activity. 40 The LRC-EC system is especially useful for multi-electrochemical detection of ES cell differentiation. 24,26,[29][30][31] Furthermore, the present LRC-EC system is superior to our previous devices in terms of the sensitivity due to the efficient redox cycling in the nanocavities.…”

Section: Electrochemical Imaging Of the Alp Activity In The Ebsmentioning

confidence: 99%

A local redox cycling-based electrochemical chip device with nanocavities for multi-electrochemical evaluation of embryoid bodies

et al. 2015

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An electrochemical device, which consists of electrode arrays, nanocavities, and microwells, was developed for multi-electrochemical detection with high sensitivity. A local redox cycling-based electrochemical (LRC-EC) system was used for multi-electrochemical detection and signal amplification. The LRC-EC system consists of n(2) sensors with only 2n bonding pads for external connection. The nanocavities fabricated in the sensor microwells enable significant improvement of the signal amplification compared with the previous devices we have developed. The present device was successfully applied for evaluation of embryoid bodies (EBs) from embryonic stem (ES) cells via electrochemical measurements of alkaline phosphatase (ALP) activity in the EBs. In addition, the EBs were successfully trapped in the sensor microwells of the device using dielectrophoresis (DEP) manipulation, which led to high-throughput cell analysis. This device is considered to be useful for multi-electrochemical detection and imaging for bioassays including cell analysis.

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A Pt layer/Pt disk electrode configuration to evaluate respiration and alkaline phosphatase activities of mouse embryoid bodies

Cited by 24 publications

References 32 publications

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

Micro/nanoelectrochemical probe and chip devices for evaluation of three-dimensional cultured cells

A local redox cycling-based electrochemical chip device with nanocavities for multi-electrochemical evaluation of embryoid bodies

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