If I wanted to develop a resistance pancreatic adenocarcinoma cells, from a panc1 cell-line, against gemcitabine, and I don’t have gemcitabine as a raw active material by itself, I have it as a medication for iv infusion in the form of hyrdrochloride, containing other excipients such as Mannitol, sodium acetate, HCL/NaOH, do you know if using this medication instead of gemcitabine compound, alone, will work well or it might harm/ give false results in MTT or in resistance??
Many thanks for helping...
It is not clear, Are where cells in vitro or in vivo? Usually metabolites of medicine substance are not so active or not so stable.
In my opinion, the use of such a medical formulation instead of the pure drug itself is basically not a problem.
In our lab, we did this too (sometimes) and did not encounter any difficulties. However, you have to be aware of possible effects caused by the accessory ingredients of your formulation (sterility? pH? ...). From my point of view, the substances you mentioned won't be much of a problem, given the dilutions you use in your experiments are sufficiently high for all compounds.
Good luck.
Based on your other ingredients, mannitol and acetate are not toxic at reasonable concentrations. Mannitol can affect cell osmolality, but if diluted enough should be good. Acetate at high concs can affect pH. So check pH
However, for your control you must use the same vehicle as in the original formulation, which would give you a better comparison. So your vehicle is mannitol/acetate/NaCl combination at same concentration as the drug.
If you do not know the vehicle makeup, you can use the formulation but just make sure the dilutions don't affect pH or viability.
I think it is OK to use the medical formulation. You need to make sure the concentration is around the IC50 range for the cell types you are using.
As long as you are using the drug at such concentrations, it shouldn't give any false positive results in cell function assays.
Good luck.
Fedor Vitaliyevich Donenko, Thank you. Its is in vitro , panc1 cell-line.
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