DMSO Usage in Cell Culture
See Also
The Truth About DMSO in Cancer Treatment: Unproven or Disproven | Ultimate in Integrative Holistic Cancer Therapy, Houston, TexasDIMETHYLSULFOXIDE (DMSO): Overview, Uses, Side Effects, Precautions, Interactions, Dosing and ReviewsDmso (dimethylsulfoxide): Health Benefits, Side Effects, Uses, Dose & PrecautionsDMSO: Uses and RisksDimethyl sulfoxide (DMSO) is an organic compound with the formula of (CH3)2SO. DMSO is frequently used in cell banking applications as a cryoprotectant. DMSO prevents intracellular and extracellular crystals from forming in cells during the freezing process. For most cryopreservation applications, DMSO is used at 10% concentration and is usually combined with saline or serum albumin.
Hydrophobic peptides can be easily dissolved in DMSO. However, peptides in DMSO could be cytotoxic to the cells although DMSO increases cell permeability. A high concentration of DMSO should never be used for cell culture. 5% is very high and will be dissolving the cell membranes. Most cell lines can tolerate 0.5% DMSO and some cells can tolerate up to 1% without severe cytotoxicity. However, primary cell cultures are far more sensitive. So if it is the primary cell you are using then do a further dose/response curve (viability) at concentrations below 0.1%.
So for very hydrophobic peptides, try to dissolve the peptide in a small amount of DMSO (30-50ul, 100%), and then slowly drop the solution to a stirred aqueous buffer solution like PBS or your desired buffer to the desired concentration. If the resulting peptide solution begins to show turbidity, you have reached the limit of solubility. Sonication will help to dissolve the peptides.
Rule of thumb:
- 0.1% DMSO is considered to be safe for almost all cells.
- 0.5% DMSO as the final concentration has been used widely for cell culture without cytotoxicity.
- 1% DMSO doesn't cause any toxicity to some cells but 0.5% DMSO is recommended.
- 5% DMSO was used successfully for some cells.
To keep the final concentration to 0.5%, you can make 200x stock in 100% DMSO.
FAQs
Rule of thumb:
What percentage of DMSO is needed for cell culture? ›
The sensitivity to DMSO can vary on two log magnitudes between various cell lines (normal and cancerous). Usually, it is better not to go higher than 0.1% DMSO (with two controls: the first one without drug and without 0.1% DMSO) and the second one without drug and with 0.1%DMSO).
Why use DMSO in cell culture? ›
As a cryoprotective agent, DMSO is added to cell freezing medium to prevent the formation of ice crystals during the freezing process, otherwise cells would be destroyed and dead. It is commonly used in cell banking. DMSO has a low toxicity to cells.
Is 10% DMSO toxic to cells? ›
Results: The results showed that DMSO had a significant toxicity and inhibition of proliferation on 4 cell lines, at the concentrations of 10%, 5%, 2.5% and 1.25% (p<0.0001). Methanol and ethanol inhibited cellular proliferation at the concentrations of 10% and 5% (p<0.0001).
How much DMSO should I use? ›
For nerve pain: 50% DMSO solution has been used 4 times daily for up to 3 weeks. For osteoarthritis: 25% DMSO gel has been used 3 times a day, and 45.5% DMSO topical solution has been used 4 times a day.
Can cells survive in 1% DMSO? ›
Most cell lines can tolerate 0.5% DMSO and some cells can tolerate up to 1% without severe cytotoxicity. However, primary cell cultures are far more sensitive.
Can you use too much DMSO? ›
DMSO can also cause a deadly reaction when used in high concentrations. Using DMSO by mouth can cause: Dizziness. Drowsiness.
How to dilute DMSO for cell culture? ›
Dilution for Cell Culture:
To do this, add the DMSO-peptide solution dropwise to sterile PBS or cell culture medium while gently mixing. Continue mixing until the DMSO-peptide solution is well-diluted and hom*ogenous.
What grade DMSO for cell culture? ›
T6316. High grade for peristaltic pumps and cell culture, I.D.
Does DMSO need to be sterilized for cell culture? ›
DMSO, Sterile Filtered is prepared by filter sterilizing DMSO through 0.2 micron filter. DMSO, Sterile Filtered is suitable for cell culture and tissue culture work that require elimination of all potential source of contamination.
DMSO, like many other organic solvents, has the ability to inhibit the growth of bacteria and/or kill them.
Why is DMSO controversial? ›
During clinical trials in patients in the mid-1960s, DMSO was reported to be effective against such conditions as arthritis, gout and various infections, but toxic- ity studies in laboratory animals done around the same time disclosed the DMSO could cause changes in the lens of the eye.
Why use 10% DMSO? ›
Cell cultures for research are often stored in liquid nitrogen using slow cooling methodology. To prevent damage by intracellular ice crystals, cells are slowly cooled to −80 degrees in the presence of 10% DMSO before storage in liquid nitrogen3.
What are the disadvantages of using DMSO? ›
Dimethyl sulfoxide may cause you to have a garlic-like taste within a few minutes after the medicine is put into the bladder. This effect may last for several hours. It may also cause your breath and skin to have a garlic-like odor, which may last up to 72 hours.
Should I use 5 or 10% DMSO for freezing cells? ›
5-10% DMSO is usually used for freezing cells. DMSO prevents the formation of ice crystals which otherwise lyses the cells during thawing. using a low concentration of DMSO means cell may not recover well while Thawing, because of ice crystal formation.
What percentage of DMSO is toxic to bacteria? ›
Apparently, the widely used biological solvent DMSO was toxic to microorganisms at high concentration, and the concentration should be less than 2% when using DMSO as a solvent.
What concentration of DMSO is used in cell storage? ›
DMSO is typically added to the media used for freezing cells at a concentration ranging from 5–10% to protect them from the harsh conditions. Cell banking typically uses 2 mL vials that are filled via pipette in a relatively quick process.
