What s the maximum allowable limit of DMSO as a solvent in cell culture - (Sep/29/2006 )

Hai,

This I feel is a very good forum to interact.My question is to know the maximum allowable percentage of DMSO as a solvent in cell culture.Some literature says its upto 0.1%.Is that true .then I wonder how 10% of it is functions well in the freezing medium without much cytotoxicity.Can anybody explain the magic behind that

---

Hi!
I have done experiments with DMSO up to 0.2%, but I guess it depends on the cell line you use (I was working with HL-60, Jurkat and MOLT-4). This is the concentration cells can bear and keep on living.
During freezing process DMSO helps you to avoid that H2O forms crystals into cells, and since crystals are bigger in volume, cell would "explode".
I have been told to be very fast in freezing an defrosting cells so that cells aren't in contact too much time with such a high concentration of DMSO.
Hope this is right and clear!

---

---

Thanks for the reply.I understand long term exposure to DMSO in liquid form is the culprit. Now i am doing experiments by keeping DMSO upto 0.5%.I have to do an analysis with different concentration of DMSO in those cell lines with which I am working. I will come back later with the inference.

---

Hi all,

I am stucking with one problem and hope people have answer to this.

i have to study the chemopreventive effect of kaempferol on cancer cell line. i dissolved it in DMSO

(100%) and as i dilute it in DMEM( without FCS), there was clotting and turbidity.

Now suggest me what do i do? Do i use antother suitable solvent, if you people have any idea or suggestion then please help me.


waiting for your kind reply.

---

using up to 0.5% may work but as others suggested you have to find out the maximal tolerable conc which depends on your kind of cells; in any case, one has to run control experiments with DMSO alone

DMSO in freeze buffer prevents crystalization of water which would damage cells; it is cytotoxic and should only be in contact during freezing and, later, thawing, and then rapidly removed; during freezed storage, DMSO seems to do no harm

---

I was always told freeze slow - thaw fast

---

Dear Anand,

In my experience the maximum concentration for cell based assays for drug discovery is 0.1%. I have used both primary (chondrocytes & endothelial cells) and cell lines (caco-2, J774, Hela's, THP-1 etc) in drug discovery experiments. The problem you have sometimes is that the chemists produce highly insoluble compounds that can only be dissolved in solvents like DMSO/ethanol and not in aqueous solution. Remember to always run a solvent control in your experimental protocol.
However in cryopreservation of cells, a working concentration of 10% is used. The DMSO stops/protects the cell membrane from the freezing process i.e. ice crystal formation/puncturing of cell membrane. If however the cells are left too long in this concentration, then the cells rapidly lose viability.


Hope this answers some of your questions

Kindest regards

Rhombus

---

I've been dealing with this exact situation recently. I finally had to conduct a titration of DMSO alone from 0.1% to 5% of total volume to determine how much I could use before function became altered. What suprised me was that even at 0.1% I could detect a slight decrease in function, and this began to get significant at 0.5%. Take home message... use as little as possible, and always have a solvent control.

---