Using live-cell imaging in cell counting — The cytotoxicity of DMSO

Longer DMSO processing times result in greater impacts

DMSO was added after seeding Hep G2 cells in a six-well cell culture plate at a density of 2.5 x 10⁵ cells per well. Live-cell imaging was used to observe DMSO cytotoxicity.

The day following seeding, the growth medium of each test well was replaced with a medium solution containing DMSO at concentration levels of 0.1 percent, 0.5 percent, 1 percent, 3 percent, and 5 percent, respectively. The culture plate was then set in a BioStudio-T* device inside a CO₂ incubator and cultured for 72 hours, during which phase-contrast images were captured once every 12 hours. A CA module was then used to perform image analysis on the phase-contrast images to calculate the cell occupied area ratio.*This product has been discontinued. If you have any questions about cell cultures, assays, or products, please contact us using the contact form.

The results reveal that increased concentrations of added DMSO correspondingly slowed the growth rate of cell confluency. It can be seen that there was no cell proliferation at a DMSO concentration of 5 percent.

The confluency of each test well was calculated as a ratio of the control well (the DMSO-free well) and graphed with the DMSO concentration values plotted on the horizontal axis. It can be seen that cell proliferation is inhibited over time following the addition of DMSO. Growth is particularly inhibited at concentration levels of 3 percent and 5 percent, illustrating the cytotoxicity of DMSO.

Continuous measurement of cell proliferation when growing cultures with added DMSO is now possible, which demonstrates the effectiveness of this analysis method in evaluating various measures following the addition of a drug, such as monitoring how long the drug effects last and identifying when those effects first appear.