Hi researchers!
I have gained something strange in the result of mtt assay of the PBMC.
I was testing an herbal extract that is declared to be safe for normal blood cells by researchers but I saw that ODs of the different doses on PBMc such as Ic50/4 , IC50/2 ,IC50 and 2 IC50 were the same!and all of them were half of the negative control!!!
Can any one interpret that for me?
I have two assumption:
1: The PBMCs behave differently. Theoreticaly, these should not be confused with blood cells. Check the literature that how this drug behaves with monocytes and dendritic cells.
2: the extract possibly contains some solvent that may be interfering with the result. Try running MTT against the solvent as well.
Also, there is an acceptable limit or range of the viability. sometimes MTT assay is not dose dependent. Increasing the concentration does not pose any visible changes. Also, try to find the accurate OD wavelength, it also varies with protocols sometimes. The cells are not adequately seeded, might be another reason for fluctuations.
It might be an issue with conditions under which the cells were cultured or issues with cell integrity.
Before the experiment, check the cells under the inverted microscope for confluency and morphological changes which will give you an idea if the cells are fine prior to addition of your extract.
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