I have a student who wants to look at the effects of lycopene on HeLa cells. We are trying to determine the best way to introduce it into the cell culture media. Yesterday we mixed it in DMSO, total concentration of DMSO in media about 1%, but we are concerned this will be too toxic. I have read about water-dispersible lycopene beadlets (10% lycopene) and placebo beadlets were a gift from Hoffman-La Roche (Nutley, NJ). Or forming some type of micell to introduce it? Any thoughts would be great!
I would like to suggest that you decrease the final concentration of DMSO to 0.5% to minimize the solvent effect. In addition, the use of water-dispersible lycopene beadlets (10% lycopene) and placebo beadlets should be better than using DMSO to increase the solubility of lycopene. After addition to the cell culture medium the solubility of lycopene will be dramatically changed and you will have difficulties to determine the real concentration of lycopene that is available to the cultured cells.
Hi Steve; As far as DMSO goes it would be better to try for 0.01% or lower in the medium. Also human serum, at least, contains lycopene and other carotinoids. I assume that you are using FBS. You may want to check what it contains already in terms of cartinoids. The Mayo Clinic has a nice monograph on lycopene in humans with some information on concentrations, etc. which may be helpful. Also since lycopene is fat soluble (oil) you may be able to go that route instead of the toxic solvent way.
In our hands human or mouse keratinocytes and respective cell lines such as HaCaT cells tolerated well up to 1% DMSO (for adding retinoic acid), though in those cases epidermal differentiation might have been enhanced by DMSO (in the old days DMSO had been considered as one of the 'inducers'). Thus, routinely we kept DMSO up to or below 0.5% as already suggested by Yau-Huei Wei.
Thanks for all the help! Looked at the cells today, one of the control plates with DMSO and no Lycopene actually looked reasonable, not as much growth as the control with no DMSO but better than I had expected. The interesting plates were the ones with 50 and 100 micro-molar concentrations of Lycopene. Lots of floating cells and the attached cells were full of granules, which I am assuming is the Lycopene or its metabolites, Maybe a combined effect of the high Lycopene with the DMSO? We are letting these cultures grow out to see how they develop with just normal media changes, no DMSO or Lycopene. Is the anyway to add pictures to this forum?
We just performed cell counts and those treated with just 1% DMSO and those with the lycopene were all the same statistically and they were are significant from the control with only media. So we are going back and trying dishes with 0, .1, .5 and 1% DMSO since this is the only option we have at this time....
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