I have performed MTT assay of Hydrophobic peptides against MDA-MB hrs and -231 cell line at Passage no P12 and P13 at 3 different time points, 24 hrs, 48 hrs and 72 hrs. At 48 hrs, 10 uM concentration of thedrug is showing approximately 70% of cell viability in P12 passage whereas in P13, the same concentration shows approxiamately 60% of cell viability. What could be the possible explanation? Can the Hydrophobicity of the drug may cause any effect to the cell viabiity?
I can't advise on the specific reason, however, in general cells become more senescent with increasing passage number, so the P13 cells may be more susceptible to the drug.
Hello Kasturee,
In my opinion, P12 and P13 is too much old cells, at the time of every passage, may be cells getting changed their physio-morphological changes due to sort of stress or starvation during sub-culturing, incubation etc.
I think the results you observed may be due to kind of changes in cell physiology or may be your sample in not homogeneous when you applied.
In my opinion try to treat sample in early passage of cells and even you get same results then just verify your sample (all ready used one and freshly prepared one).
I don't have specific reason for your question but i think this information may contribute to overcome from your situation.
All best wishes for your endeavor.
Hi Kasturee:
(1) To me, 60% vs 70% viability by MTT is mild difference, many factors can cause this: such as minor difference in cell density; growth status; or shaking-hands, bla bla.
(2) If this result is very repeatable by MTT, it would be interesting to validate it by another cell viability assay, such as PI staining and look it on FACS, because MTT assay depend on activity of oxidoreductase enzymes, and PI assay associate with permeability of cell membrane. If both results are repeatable and consistent, you can draw the conclusion that different passage times affect cell viability.
(3) To verify whether the drug cause the cell viability change, you can compare your drug treated groups with your solvent groups in the same cell.
Regards
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