DMSO Usage in Cell Culture
Dimethyl 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
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.
What concentration of DMSO kills cells? ›
Consistent with previous findings on other cell types, DMSO at concentrations ≥ 1% reduced cell viability and accelerated cellular damage by altering mitochondrial membrane potential and increasing ROS in our model.
Does DMSO increase cell growth? ›
It can be seen that cell proliferation is inhibited over time following the addition of DMSO. Growth is particularly inhibited at concentration levels of 3 percent and 5 percent, illustrating the cytotoxicity of DMSO.
Why did you need to remove DMSO from the cell media? ›
Although the cell products have demonstrated excellent post thaw viability, DMSO is considered a potential cause of infusion related adverse events. 1,2 Therefore, removal of DMSO and cell lysis products by washing the cell product after thawing may reduce the severity of some transplant related complications.
Why use DMSO as a control? ›
Answer and Explanation: Dimethyl sulfoxide (DMSO) is used as a control because it serves as a medium through which a certain chemical is delivered. This helps the scientist determine if any changes he is observing are due to the DMSO or if they are due to the chemical he or she is testing.
How much DMSO to use in cell culture? ›
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.
Can cells survive in 1% DMSO? ›
Higher DMSO concentration (1.0%) and higher cell density (2 × 107 per milliliter) also significantly decreased cell survival to 73%.
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 are the cons of DMSO? ›
DMSO use has been associated with the following potential side effects: skin rash, erythema, itchiness, scaly skin, skin thickening, hives, blistering, skin pain, nausea, diarrhea, headache, garlic-like taste in mouth, garlic-like breath, garlic-like body odor, transient hemolysis, dizziness, sedation, kidney injury, ...
Why is DMSO controversial? ›
The biggest concern of DMSO as a solvent is that when it gets on the skin it will cause anything on the skin to be absorbed.
Freezing down cells in DMSO. The use of a freezing medium should contain a cryoprotective agent like dimethyl sulfoxide (DMSO) or glycerol. These agents reduce culture-medium freezing points and allow for slower cooling rates; greatly reducing the risk of ice crystal formation which can damage and kill untreated cells.
What is the purpose of using DMSO? ›
DMSO is used topically to decrease pain and speed the healing of wounds, burns, and muscle and skeletal injuries. DMSO is also used topically to treat painful conditions such as headache, inflammation, osteoarthritis, rheumatoid arthritis, and severe facial pain called tic douloureux.
What is filter DMSO 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.
Why is DMSO so good? ›
Dimethylsulfoxide (DMSO) has been recommended as a treatment for endotoxemia and SIRS because of its antiinflammatory and reactive oxygen species (ROS)–scavenging benefits.
How does DMSO damage cells? ›
The suggested mechanism for DMSO cytotoxicity is the effect on the physical properties of the phospholipids in membranes. As an amphipathic solvent, DMSO can interact with the plasma membrane allowing pores formation, which contribute to decrease membrane selectivity and increases cell permeability [14].
Why is DMSO not FDA approved? ›
It was even actively investigated for medical use, but because of adverse effects especially affecting the eyes, most clinical trials were halted in 1965 by the United States Food and Drug Administration (FDA). At this point in time, DMSO was seen as extremely toxic, comparable to thalidomide4,5.
Should I use 5 or 10% DMSO for freezing cells? ›
To avoid damage to the cell during freezing, a cryoprotectant is added to the growth medium in which the cells are to be frozen. Glycerol or DMSO (dimethyl sulphoxide) in 10% concentration is most commonly used.
Is 5% DMSO toxic? ›
Toxicity became significant at 0.1% DMSO, as it reached approximately 5–12%. Concentrations above 0.1% should not be used, as significant toxicity was observed (≈15%).
Is 10% DMSO toxic to mice? ›
Less than 10 % DMSO should not be toxic to the mice.
What is the acceptable range of DMSO? ›
Available data indicate that they are less toxic in acute or short-term studies and negative in genotoxicity studies. It is considered that amounts of these residual solvents of 50 mg per day or less (corresponding to 5,000 ppm or 0.5 percent under Option 1) would be acceptable without justification.
