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

Abstract

Oxygen consumption rate (OCR) of mouse embryoid body (EB) was measured repeatedly in glucose-depleted medium or PBS solution. Non-invasive nature of the scanning electrochemical microscopy allowed sequential OCR measurement of the identical EB sample for more than 24 h. We found that the OCR value for the undifferentiated EB (cultured for 2 days) reached almost zero after 12 h exposure in PBS. On the contrary, the OCR value for the differentiated EB (cultured for 8 days) indicated 40 to 28% normal to the OCR at 0 h-incubation after 24 h exposure in PBS-based solution. It is suggested that differentiated cells indicated a stronger adaptation to glucose deprivation comparing to the undifferentiated cells.

Scanning Probe Microscopy for Nanoscale Electrochemical Imaging

“…The respiratory activity of live HeLa microtissues has been assessed by monitoring the oxygen reduction current in constant height mode SECM. We reported SECM investigation of the respiration activity of mouse embryonic stem cells, , rat primary hepatocyte spheroids, muscle cells embedded in a hydrogel-metallic glass wire complex, the β-galactosidase activity of cellular aggregates as a biomarker for senescence, the alkaline phosphatase activity of single cells entrapped in microwells with ring-electrode arrays, and intracellular enzymatic activities . Membrane protein expression levels have also been evaluated using SECM. , …”

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

“…16 The effects of glucose-depleted media on O 2 consumption of EBs were investigated using SECM. 17 According to the results, the differentiated cells showed a stronger adaptation to glucose deprivation than the undifferentiated cells. Furthermore, the relationship between respiratory activity and gene expression in EBs was investigated.…”

Bioanalytical chemistry with scanning electrochemical microscopy

“…In particular, the recent development of nanoelectrode fabrication has made it possible to analyze a single cell at higher spatio‐temporal resolutions 67 . Examples include high‐resolution imaging of biological samples, 68–70 measurement of oxygen consumption, 71–80 monitoring of ROS/RNS inside and outside cells, 81–89 and monitoring of exocytosis 90–94 . Because SECM bioanalysis with nanoelectrodes has good sensitivity, signal to noise ratio, and non‐invasive in nature, it is suitable for detecting small amounts of chemical species in a single cell.…”