Hi! I'm currently working on establishing an in-vitro model for diabetes-research concerning DNA-methylation, and therefore I'm cultivating HepG2 and HEK293 cells at different glucose concentrations varying from 5.6 mM to 30 mM. When measuring the cell viability using TrypanBlue I could observe a slight decrease the higher the glucose concentrations was, however measuring the cell viability using alamarBlue the viability increased. So I was wondering if the high glucose in the medium could have any effect on the outcome of the alamarBlue assay? Could anyone help me? I'm kind of confused by my results. Thank you in advance for any kind of tips and answers :)
Are you using media-only blank controls for your Alamar blue assays? That will answer your question about the glucose as well as provide the correct baseline from which to assess viability.
If I recall it correctly, the Alamar blue assay is actually measuring mitochondrial metabolic turnover and is thus regarded as a viability assay. In theory, higher glucose levels can lead to a higher metabolic activity which might be misinterpreted as increased viability.
10.1016/j.gene.2015.08.009
Check out the given publication which disuses the shortcomings of MTT-based viability assays which have a similar mechanism as AB.
Best
S.
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