MTT vs. Live/Dead assays - Viability assay differences (May/30/2007 )
Hi,
I was trying to assess viability of human MSCs and wasn't sure what the practical difference between the MTT assay and Live/Dead assay (molecular probes) is. I know how they work, but is one used more often than the other in some cases or is one more sensitive than the other. Since the MTT measures mitochondrial activity and the Live/Dead measures nuclear envelope integrity, which of the above events would happen first/more readily during cell death? Thanks for your help
bioegrad
I was trying to assess viability of human MSCs and wasn't sure what the practical difference between the MTT assay and Live/Dead assay (molecular probes) is. I know how they work, but is one used more often than the other in some cases or is one more sensitive than the other. Since the MTT measures mitochondrial activity and the Live/Dead measures nuclear envelope integrity, which of the above events would happen first/more readily during cell death? Thanks for your help
bioegrad
the MTT is not to use for discrimination between apoptosis and necrosis; I do not know exactly the Live/dead assay of MP as they offer varius assays but nuclear envelope desintegration is more more a matter of apoptosis; nevertehless, it is always a good thing to test cytotoxicity with various assays, especially to differ between necrosis and apoptosis...
ya, MTT cannot differentiate necrosis and apoptosis.
By the way, you want to check the cells viability after you treat the cells with?
In my experiment done before, i use some bacteria protein (including toxin in batcreia) to treat the cells. after that i used XTT from Sigma (a similar product like MTT, but the final substrate is already in soluble form), the reading is incorrect!!! the more cells die, the reading is higher (should be lower)!!! after some time only i realised the some protein that i used to treat the cells can react to XTT and the more protein i put the higher my reading is...
So, be careful with what you treat your cells with.
If it really happen, you can try treat your cells for (few hrs), wash away the chemicals and put in MTT to incubate.