I'm working with the SH-SY5Y cell line and I need to treat them with rutin. The best way I have found to solubilize this flavonoid is with DMSO 100%, but since it's toxic for the cells, I've decided to dilute the stock solution using PBS until 5mM (DMSO 1%).
My plan is to adjust the pH, filter it and then proceed to the next dilutions before reaching the final concentrations (with DMSO 0.01%) to use in the cells because this way I'll ensure the effects observed are from rutin, not the pH. However, a colleague told me I should not adjust the pH - it could even be a bad thing.
I'm confused. Should I adjust the pH after all?
There is no need to adjust the pH in cell culture, DMSO at low concentrations is not considered cytotoxic, we use it at a higher concentration of 10% for freezing cells at -80 routinely to keep cells in the freezing stage and it is also used to dissolve some active substances in low concentrations
Adjusting the pH may require you to add (NaOH) or (HCl), and these undoubtedly have effects on the cells and their vital processes.
Thank you for your answer! I just wonder if the effects I'll observe in the cells will be due to rutin or due to the difference in pH. When I prepare the 5mM solution (DMSO 1%) using PBS, I get pH = 7. I know the cells should be kept in pH = 7,4. I'm worried about that...
DMSO should not affect the pH of your solutions. In fact, so long as you are using cell culture grade sterile DMSO, there is no need for you to filter it, either. Just dilute and go. Not only is the dilution so tiny as to be inconsequential, your medium has buffering capacity.
Thank you! It makes sense... But what about rutin? Does it affect pH? I'm also working with ayahuasca and it does change the pH. I have to ajust it before using it in the cells. But I dilute it in PBS, there is no DMSO involved.
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