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DMSO and cellular culture


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#1 eliane

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Posted 22 October 2009 - 11:22 AM

Hello

I have the following problem: I have to use a compound that dissolves only in DMSO 100%. I need to use it in cellular cultures of certain endothelial cells and stellate cells, that are exquisivitely sensitive to DMSO. At 0.01% DMSO, those cells die. I need to use the compound at 1 to 10 micromol/l concentrations, and I am not able to make a stock solution above 10 mmol/l, or this compound does not dissolve even in DMSO. So I can't go below 0.01% concentration of DMSO in my cells (am I clear or not at all ???)
has anybody an idea ?

Thanks a lot +++

#2 jah

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Posted 22 October 2009 - 12:10 PM

How about another solvent - Ethanol, methanol or acetone? You might also consider a 'carrier' molecule. 2-hydroxypropyl-beta-cyclodextrin has been used with several hydrophobic small molecules

#3 jakatta70

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Posted 22 October 2009 - 02:10 PM

Hello

I have the following problem: I have to use a compound that dissolves only in DMSO 100%. I need to use it in cellular cultures of certain endothelial cells and stellate cells, that are exquisivitely sensitive to DMSO. At 0.01% DMSO, those cells die. I need to use the compound at 1 to 10 micromol/l concentrations, and I am not able to make a stock solution above 10 mmol/l, or this compound does not dissolve even in DMSO. So I can't go below 0.01% concentration of DMSO in my cells (am I clear or not at all ???)
has anybody an idea ?

Thanks a lot +++


Hi,

I experienced a similar problem recently. I performed a concentration range of DMSO with my Caco-2 cells and assessed for cell death via an LDH assay. I found that concentrations of 1%+ were toxic. So in order to get rounf this I made a 10 mM stock solution of my drug in 100% DMSO. I want t a final concentration of 10 uM that the cells are exposed to in 1ml of media. This equals 1 ul of stock solution to 1 ml of media which represents 0.1% DMSO.

Have you performed a cell death assay to confirm that your cell lines cant take anymore than 0.01% DMSO? Its just that the figure seems quite small to cause toxicity.

Perhaps make your stcok solution as concentrated as possible in 100% DMSO and then bring the concentration down by diluting in sterile water. Your drug should still remain in solution while reducing the final DMSO concentration that your cells are exposed to.

#4 bob1

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Posted 22 October 2009 - 04:27 PM

Have you performed a cell death assay to confirm that your cell lines cant take anymore than 0.01% DMSO? Its just that the figure seems quite small to cause toxicity.

The toxic effects of DMSO can be seen in most cell lines at around 0.1% (I'm not talking about death, but about altered stress gene expression etc.), ideally you need to evaluate these genes and see if they affect your system.

I have the following problem: I have to use a compound that dissolves only in DMSO 100%. I need to use it in cellular cultures of certain endothelial cells and stellate cells, that are exquisivitely sensitive to DMSO. At 0.01% DMSO, those cells die. I need to use the compound at 1 to 10 micromol/l concentrations, and I am not able to make a stock solution above 10 mmol/l, or this compound does not dissolve even in DMSO. So I can't go below 0.01% concentration of DMSO in my cells (am I clear or not at all ???)
has anybody an idea ?

Most drugs that dissolve in DMSO will also dissolve in dimethylformamide (DMF, sometimes known as the universal solvent) at a higher concentration. Perhaps you could try DMF instead.

#5 eliane

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Posted 23 October 2009 - 12:51 PM

How about another solvent - Ethanol, methanol or acetone? You might also consider a 'carrier' molecule. 2-hydroxypropyl-beta-cyclodextrin has been used with several hydrophobic small molecules

thanks I will try that and tell you

#6 eliane

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Posted 23 October 2009 - 12:54 PM

hi !

thanks for your answer

I did not perform any cell assay, but cells change their shape and detach from the well, and I can't use them for WB at all. So they seemed pretty dead. With other cells I tried the toxicity appears at 0.1%.

I will tell you when I solve the problem !

#7 eliane

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Posted 23 October 2009 - 12:56 PM

Hello

I have the following problem: I have to use a compound that dissolves only in DMSO 100%. I need to use it in cellular cultures of certain endothelial cells and stellate cells, that are exquisivitely sensitive to DMSO. At 0.01% DMSO, those cells die. I need to use the compound at 1 to 10 micromol/l concentrations, and I am not able to make a stock solution above 10 mmol/l, or this compound does not dissolve even in DMSO. So I can't go below 0.01% concentration of DMSO in my cells (am I clear or not at all ???)
has anybody an idea ?

Thanks a lot +++


Hi,

I experienced a similar problem recently. I performed a concentration range of DMSO with my Caco-2 cells and assessed for cell death via an LDH assay. I found that concentrations of 1%+ were toxic. So in order to get rounf this I made a 10 mM stock solution of my drug in 100% DMSO. I want t a final concentration of 10 uM that the cells are exposed to in 1ml of media. This equals 1 ul of stock solution to 1 ml of media which represents 0.1% DMSO.

Have you performed a cell death assay to confirm that your cell lines cant take anymore than 0.01% DMSO? Its just that the figure seems quite small to cause toxicity.

Perhaps make your stcok solution as concentrated as possible in 100% DMSO and then bring the concentration down by diluting in sterile water. Your drug should still remain in solution while reducing the final DMSO concentration that your cells are exposed to.


And I forgot to add that as I want to reach concentrations of 10 micromol/l, and my higher stock solution is 10 mmol/l, I can't go below 0.1% DMSO ...




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