Cancer cells were treated with conditioned media. In the MTT curves it shows inhibitory effect while cells treated in AlamarBlue are proliferating faster (the same cell line, the same conditioned medium, the same day experiment).
Very strange result. In obvious sense, results of both assays should be correlated. Did you put equal numbers of cells per well for both assays? I ask you about it, because Alamar Blue assay is much more sensitive and obviously requires to add fewer number of cells per well. It may be, you saw inhibitory effect due to overgrowing of cells assigned to determine MTT transformation. (??)
I think so, number Cell per well is very important for exelelent result.
Did you ever figure this out? We have noticed this exact phenomenon as well but it may be cell type specific. We see contradictory results (increased growth alamar blue, decreased growth MTT) in HT-29 cells but not other cell types. Based on the visual inspection of the cells, the MTT data appears to be correct in those cells. We also looked at absorbance and fluorescence for Alamar Blue and at least those are in agreement as far as magnitude of effects. We also looked at multiple time points with Alamar Blue (every 30 minutes for 3 hours) and the result is the same. Very interested to hear if you have any new thoughts on this.
Some compounds are nasty with the MTT (or MTS) assays ... and it can indeed be cell type-dependent.
See the attached articles.
Best regards
Robert
I am eager to know the answer, too. I learn that MTT assay relies on mitochondrial oxidative phosphorylation while AlamarBlue relies on reducing substance produced by cell metabolism. Is it possible that they are different in using substrate to convert the chromophore?
- Badges
- Science topic
- Similar topics
- Geoscience
- Asia
- India