Dear all,
I recently encountered a problem with how to add exogenous ceramide to cell culture to monitor their biological effects. Currently, we want to add two ceramides to the cell culture： Ceramide (d18:0/24:0) and Glucosylceramide (plant), but we found it was only a little soluble in chloroform or DMF. These organic solvents cannot add 1% to cell culture which will also limit these ceramides' delivery to cells. Does have any better buffer that could form the lipid vesicle or stable suspension for subsequent cell treatment? Please the lipids experts or biologists give us some suggestions or advice, thank you very much!
Dear Dr. Bingpeng Yan
A solvent mixture of ethanol and dodecane in the ratio (98:2, v/v) is able to disperse natural ceramide into aqueous solution.
You may want to refer to the paper attached below.
https://febs.onlinelibrary.wiley.com/doi/abs/10.1016/0014-5793%2894%2901428-4?sid=nlm%3Apubmed
Refer also to the link below. See Method D: Solubilization using ethanol: dodecane (98:2,v/v) for use in cell cultures
https://www.caymanchem.com/news/Sphingolipid-Advice#:~:text=Dissolve%20ceramide%20(from%20bovine%20brain,with%20the%20medium%20using%20agitation.&text=Transfer%20to%20culture%20dishes.
Regards,
Malcolm Nobre
Dear Malcolm,
Many thanks for your detailed answer, it's quite useful for my experiments.
However, the dodecane isn't on our hands, may I know if this one could be replaced by Hexane? In addition, if we used the ethanol: dodecane (98:2,v/v) solvent to dissolve ceramide, do you have any suggested reference for the around maximum solubility?
Thank you very much!
Best,
Bingpeng
Malcolm Nobre
I borrowed the dodecane from my chemical friends but they all never use this one. So may I know if could we use the ethanol: hexane (98:2,v/v) solvent instead?
Thank you very much!
Look forward to hearing you back soon.
Best,
Bingpeng
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