I need to do some tests in cell culture with extracts oils (rosehip) and would like to know if anyone has any protocol to solubilize in DMSO.
Hola Marisol,
If your extracts are poorly soluble in DMSO, then you will not be able to prepare concentrated stocks. You can try to sonicate your samples or heat them up to 37 ºC (but not at higher temperatures) in order to avoid any possible thermal degradation of the secondary metabolites present in your extracts.
However, in such cases you will have to decide between employing the standard DMSO-based bioassay or, alternatively, preparing a highly concentrated stock in a different, more appropriate solvent for your extracts. I would recommend to evaluate first the effect of these alternative solvents on your cell lines before making a decission.
¡Saludos!
You may follow other protocols that have been used by people to solubilize other essential oils to use them in cell culture. Follow the methods available in the literature.
Non-essential oils are usually not soluble in DMSO and later in the medium. Try other solvent such as ethanol, use carriers such as cyclodextrins or albumins (BSA, FBS), or use biocompatible emulsifiers. You can buy expensive proprietary solubilizing agents. Do not forget to control for any of these. The oils might separate when diluting into the medium (milky clouds) or even build greasy drops. Undissolved lipids are highly undesirable. The added lipophilic compounds will be adsorbed by the plastics, shuttled around by medium constituents (e.g. albumins from BSA), and will typically be quickly absorbed by the cells.
Never use DMSO as a solvent in cell culture. From my personal experience (and from other people's) DMSO interact with a lot of cell functions and you will get completely unreliable data. Ethanol is OK but only with a max final concentration of 0.01%. I used to complex not water soluble compounds (in my case long chain fatty acids) in FBS by warming and putting them in a sonicating water bath. It worked very well, the micelles formed (there is an external layer of Fetuin A that surrounds your compounds and it is able even to put in solution completely insoluble inorganic salts, such as hydroxyapatite) are very stable and they are readily up-taken by the cells. You can try also other complexing agents but if FBS work it could be better because probably you are going to add it to the culture media anyway, so you may be able to cut down a bit in the number of the controls.
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