I need some alternative dilution of tetracycline pellets instead of ethanol for in vitro studies. Ethanol is a very aggressive agent to the explants and increases the oxidation percentage in my studies.
Have you considered using dimethyl sulfoxide (DMSO)?
Try to prepare 100X or 200X tetracycline solutions in pure DMSO, then when diluting to 1X in the culture medium, the final concentration of DMSO will be 1 or 0.5% v/v and will not be harmful to the cells. Previously, you should check if the solubility of tetracycline in DMSO allows to prepare 100 or 200X solutions.
Thank you José Luis Duque-Rivera
Do you know if this concentration will not be harmful even in plant cell culture?
In plant cell cultures I do not know for sure but in human cells the tendency is to use a maximum of 0.5% v/v.
You could do a preliminary assay of three treatments with the same amount of tissue in which you evaluate some key biological variable, for example, cell viability applying the following scheme:
Treatment 1 (control A): place 1 uL of PBS (or the physiological buffer of preference) for every 199 uL of culture medium.
Treatment 2 (control B): place 1uL of pure DMSO for every 199 uL of culture medium.
Treatment 3 (tetracycline): place 1 uL of 200X tetracycline for every 199 uL of culture medium.
If the amount of DMSO is harmless, the two controls should not show statistically significant differences. Perhaps it is convenient for a statistician to advise you as to the number of replicas to be used for each treatment, in principle there should be no less than three. You could dispense with tetracycline treatment because the important thing in this phase is to establish the safety of DMSO.
Thank you José Luis Duque-Rivera . I will do some tests according to your recommendations.
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