I am dissolving some cell culture ligands in 100% DMSO to make stock solutions of 5ug/ 10ul of DMSO. Im then adding 20 ul of this stock to 1 ml of media+cells in 12 well plates to make a final concentration of 10ug/1ml cell medium. This would constitute a 2% DMSO exposure to cells per well.
when i stimulated cells with ligands diluted in DMSO and only DMSO as mock control its giving discrete pattern. Can anybody please tell me what should be the optimum percentage of DMSO exposure when it use for ligands dilutions?
Hi Nawaj,
2% is pretty high - how long are you treating? I am not sure about your cell line but in primary fibroblasts 1% DMSO shows cytotoxicity by 48 hours. Standard is to use 0.1% but some use up to 1%. This is really incubation time dependent. I always recommend erring on the side of caution, which would be as minimal as you can make it.
Good luck!
Hi Julia,
Thanks for valued reply. I will use 1%DMSO n std for various time points. Let I will reply after experiment.
2% DMSO is very high. Can you not increase the concentration of reagent in DMSO stock so that final DMSO concentration could be reduced. Also it may be good idea to include a medium control(without DMSO) just to ensure that the DMSO concentration that you are working with is not toxic..
Dear Sir,
Thanks for your valuable suggestion. yes we are now minimizing DMSO exposure 0.1 to 1% in our next experiment and also include medium control to check toxicity of DMSO.
My previous supervisor told me he checked DMSO's effects on several signal pathways when he was a postdoc. His conclusion is the maximum of DMSO concentration in cell culture is 0.1%.
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