Electrochemical Device with Interdigitated Ring Array Electrodes for Investigating the Relationship between Cardiomyocyte Differentiation from Embryonic Stem Cells and Alkaline Phosphatase Activity

Abstract

A local redox cycling-based electrochemical (LRC-EC) chip device was used to investigate the relationship between cardiomyocyte differentiation from embryonic stem (ES) cells and alkaline phosphatase (ALP) activity. In the LRC-EC chip device, ring-type interdigitated array electrodes were incorporated at n × n measurement points with only 2n bonding pads for external connection. Microwells were also fabricated at each measurement point to trap cell aggregates. To differentiate ES cells into cardiomyocytes, ES cells were three-dimensionally cultured to form simple and cystic embryoid bodies (EBs). ALP activity of these EBs was then detected using the LRC-EC chip device. The electrochemical responses for ALP activity decreased concurrently with the differentiation of ES cells into cardiomyocytes, indicating that an LRC-EC chip device is useful for evaluating cell differentiation.

Sequential Monitoring of Oxygen Consumption Rate of Mouse Embryoid Bodies in Glucose-Depleted Solution

“…The EBs at day 2 (5,000 cells/20 µL drop of medium) and the EBs at day 8 (500 cells/20 µL of medium) were prepared to obtain the undifferentiated and the differentiated EBs with similar radii, respectively. 12 Previously, we reported that the alkaline phosphatase activity of the undifferentiated EB indicated 2.8-fold larger than that of the differentiated EB. 13 As a medium, StemMedium (containing 15% fetal bovine serum (FBS), 0.8% penicillin/streptomycin, and 0.1 mM B-mercaptoethanol) was selected to prepare the EB which was used for the OCR measurement in DMEM (no glucose) (Gibco, 11966-025)-based solution.…”

Microchemistry- and MEMS-based Integrated Electrochemical Devices for Bioassay Applications

“…5). 35,38 Therefore, the proposed LRC-EC system is useful for cell screening in tissue engineering and is ideal for high-throughput biosample assays and imaging applications.…”

“…To overcome this difficulty, we proposed a novel electrochemical approach based on redox cycling, the local redox cycling-based electrochemical (LRC-EC) system. [33][34][35][36][37][38][39][40][41][42][43][44] In the LRC-EC system, two arrays of electrodes (n rows and n columns of electrodes) are orthogonally placed to produce n 2 crossing points (Fig. 4).…”

“…To address these problems, planar comb-type IDA electrodes were incorporated into the LRC-EC device, which allowed for all electrodes to be arranged on a single substrate. [34][35][36][37][38][39][40] One of the LRC-EC devices has 1,024 sensors/41 mm 2 , 39 making the device well-suited for high-throughput amperometric detection and rapid electrochemical imaging. In addition, vertically separated electrodes were incorporated into the LRC-EC device to further increase the sensor density.…”

“…The development of several types of electrochemical devices for bioassay applications, including analyses of yeast and mammalian cells, has been reported. [29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44] IDA electrodes were incorporated to amplify signals from cells and provide highly sensitive assays. Yeast cellbased IDA electrodes were utilized for the detection of hormone active chemicals.…”