If I want to perfom treatments for my stable cell line with different concentrations from any chemical compounds!
Q1)So should I seed my cells in complete culture medium or selection medium that Im still using for maintaining my cell line ?
Q2)Could I dilute my working solution directly in the media or in dist.H20 ? If so should I dilute the compound in complete medium or free serum media or medium from BOTH serum and Pen-Strep ?
Thanks in advance
Hi Yamani
1) In my experience for your stable cell line the selection medium that you are using should be continued for the propagation purposes as well as for your treatment experiments since the change in the composition of medium might confer the cells to undergo some sort of acclimitisation and in the process their overall health might be affected and eventually so does your results and their reproducibility as well.
2) If the chemical compound is soluble in DMSO then i think you should dulite it in DMSO as it is the 'gold standard' for checking the efficacy of the compounds on different cell lines. Alternatively lot of researchers do use complete media as well to dilute the compounds containing antibiotics that you have mentioned.
Alternatively to DMSO one can use ethanol, Dimethyl formamide (DMF), gamma-Butyrolactone (GBL), N-Methyl-2-pyrrolidone (NMP), Dimethylacetamide (DMAc). So out of all the available solvents you ought to refine the best solvent for your purpose. One more suggestion would be to deviate from the 'selection medium' to the 'culture medium' and see if it leads to any differences in your result.
Good Luck!!
Hello Doaa,
1) I agree with the others here that you should definitely grow the cells in the selection media.
2) Please check the solubility of the compound ( the data sheet should give you that). Storing the compounds in DMSO would give you better stability. I would always try to keep the final concentration of DMSO under 0.5% so it doesn't affect the cells. If you are using media, You could use complete media so you don't have to worry about the volume much when you add it to the cells,
I have another question
We Passage cells 3–4 times after thawing before using them in transfection experiments so is the same should happen with treatment experiments!by the way im using right now Cadmium sulfate .
I appreciate you kind response:)
Yes, that would be ideal but a couple of passages should be sufficient.
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