I'm now finding the pathway of cell death induced by drug in AGS cell lines. As my senior did cell viability assay to drug-treated cells, I didn't do it again and I use the data from her experiment. However, when I decided to find it by myself, the data I got is different from her. She found 20uM of drug can kill 40% of cells whereas I got it can kill around 60%. ( I did it 3 different times, n=3 for each time).
I'm now in half way of my research. And according to my finding, I have to restart all of my work as IC50 dose is changed.
What should I do? Please suggest me. What are the possible reasons of increased cell death of the same dose of drug?
Hi Lin, this change can be consequence of several factor (different person making the experiments, different status of the cells or different batch of the compound with different activity or concetration; although you use a commercial compound it can happen). I don't know how much work would you have to repeat but it seems you must do all of them again in order to have appropriate results uf you want to go further using other compound and comparing the results
Hope it helps
Angel
As time goes by, one major factor that changes the behavior of a cell line is the number of passages. It is always advisable to check IC50 of any previously used compound when starting work with cell lines. Of course, many other factors may affect in vitro resumts, but passage number may change significantly the course of experiments.
As say Angel and Rafael there are different factros that affect the behavviour of the cell lines. I think that one of the most important is the number of passages. By other hand several factors as different person, different status of the cells or different batch of the compound may also influence the results. Lin you must check these possibilities, further to compare the data.
Best wishes, Joan.
I think the cellular condition is the most probable reason of this inconsistancy.
I agree with Rafael, the sensitivity of a given cell line to a drug may vary according to the number of cell passage, the batch of serum (if changed), a different lot of the drug itself. It is the reason why you have to check regularly in your own hands the toxicity of your drug and infortunately repeat some of the previous experiments done, probably using a logarithmic dose response curve
Dear Zhi,
There are different cases that may lead to different results in cell viability assay including, the number of passages, precision in make the different drug concentration, cell lines ages, environmental conditions of cell lines such as incubation time, concentrations of cell lines nutrition, calibration of incubator and so on.
Best regards,
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