I am working on drug testing in C2C12 cells at myoblast and day 7 differentiated myotube stages. My drug of choice needs to be dissolved in DMSO. I have a stock solution at 8mM and to obtain a 100 uM concentration in 2ml media I need 25ul of DMSO vehicle. This leads to 1.25% V/V DMSO. I was just wondering if this will negatively impact my cells? As one replicate I did, the drug positively affected the gene expression and vehicle downregulated it. However, in second replicate all the drugs had similar levels to vehicle.
I think the DMSO concentration you are use is slightly high. I normally use 0.1- 0.5%, but up to 1% should be fine. You can treat your cells with 1.25% DMSO for the time you usually incubate your drugs and test for cell viability. Some cell lines are highly sensitive to DMSO and that might impact your data. One thing you can try is to add one extra dilution step (dilute a high DMSO concentration in medium and then dilute again to the working concentration in your wells).
Bruno is right. To exclude the effect of DMSO on cell lines and to assess correctly the action of your drugs, use the solvent at lower concentration, the best at 0,1 %. There is a high quality of DMSO 99,9% purity from Origines.
I too have seen dmso toxicity to cells.
https://www.nature.com/articles/s41598-019-40660-0
https://www.researchgate.net/post/DMSO_usage_in_cell_culture
https://www.researchgate.net/post/What_must_be_the_maximum_final_DMSO_in_a_cell_culture_plate_24_well_plate_if_I_have_to_dissolve_my_compound_in_DMSO
Article Dimethyl Sulfoxide (DMSO) Decreases Cell Proliferation and T...
a. If you need to use at the high concentratiln i would make a dmso control with the 1.25% for compairson to see how much the dmsoalone at that concentration will kills our cells with out you drug.
b. See if EtOH or acidic/basic solutions will work for your compound.
c. Try to keep things with in biological ranges for when going to animal studies the probability of success will be higher.
Say no to DMSO: dimethylsulfoxide inactivates cisplatin, carboplatin, and other platinum complexes.
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