Assessing Mitochondrial Function in Cerebral Vascular Endothelial Cells

Begin with cerebral vascular endothelial or CVE cells in culture media.

CVE cells are high energy-demanding cells that line the brain's blood vessels and are essential for brain physiology.

Place the cells into an extracellular flux cell culture plate. Incubate for cell attachment.

Replace the culture media with extracellular flux assay media that has the desired pH.

Incubate without carbon dioxide to maintain the pH.

Next, take an extracellular flux sensor cartridge, which includes sensors and a bottom plate.

Add a calibration solution to the plate and incubate without carbon dioxide to hydrate the sensors.

These sensors can measure the changes in oxygen concentration in the extracellular environment, indicative of mitochondrial activity.

Then, load the cartridge into a bioanalyzer machine and calibrate the sensors.

Replace the bottom plate with the plate containing the cells.

Measure the mitochondrial function of CVE cells.

After the appropriate number of subcultures, seed 1.6 x 10⁴ cells in 80 microliters of complete medium per well onto a 96-well extracellular flux cell culture plate and place the plate in a 37 degrees Celsius incubator with 5% carbon dioxide.

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