Strategies in developing dimethyl sulfoxide (DMSO)-free cryopreservation protocols for biotherapeutics

Advances in Cryopreservatives: Exploring Safer Alternatives

“…17 DMSO has become the "gold standard" for most cryopreservation techniques due to its ability to limit ice nucleation and promote viability after thawing. 18,19 (c) Amino acids and amines: some molecules such as proline, glutamine, betaine, L-carnitine, or tricine can be used as CPAs. 1 Some studies have demonstrated the protective effect of glutamine in the freeze thaw process.…”

“…In 1959, Lovelock et al described DMSO’s (dimethyl sulfoxide) ability to protect living cells (human and bovine red blood cells, as well as bull sperm) from freezing damage . DMSO has become the “gold standard” for most cryopreservation techniques due to its ability to limit ice nucleation and promote viability after thawing. , Amino acids and amines: some molecules such as proline, glutamine, betaine, l -carnitine, or tricine can be used as CPAs . Some studies have demonstrated the protective effect of glutamine in the freeze thaw process. , Betaine as CPA has been reported as a preservative of morphology and proliferation capacity or as a protector of cells from solute injury caused by ice formation. , Regarding l -carnitine, several studies have shown its role as a cryoprotective natural agent. , Moreover, a simple method of cryopreservation of Jurkat cells based on the use of l -proline has also been suggested, being relevant for clinical practice …”

“…In 1959, Lovelock et al described DMSO’s (dimethyl sulfoxide) ability to protect living cells (human and bovine red blood cells, as well as bull sperm) from freezing damage . DMSO has become the “gold standard” for most cryopreservation techniques due to its ability to limit ice nucleation and promote viability after thawing. , …”

“…It is important to know that dehydration will occur if there is higher osmotic pressure outside the cell. Some dehydration is beneficial for cryopreservation, as it reduces the formation of penetrating CPAs reproductive cells or even embryos of different species: avian sperm, 13 Nile tilapia sperm, 28 eel sperm, 29 Glossogobius giuris spermatozoa, 30 Mytilus embryos, 31 human oocytes, 32,33 human embryonic stem cells, 34 ovarian carcinoma cell lines, 35 and neuron-like cells 36 sulfoxides and amides DMSO, acetamide, dimethylformamide, or dimethylacetamide penetrating CPAs synthesized from dimethyl sulfide sperm cryopreservation methods, 39−53 Trichogaster fasciata embryos, 40,54 and human ovarian carcinoma cell lines (SKOV-3, OVCAR-8) 35 DMSO: limit ice nucleation and promote viability after thawing 37,38 amino acids and amines proline, glutamine, betaine, L-carnitine, or tricine neutral amino acids (β-alanine, γ-aminobutyric acid (GABA), and ε-aminocaproic acid (ε-Ahx)) proline: great osmoregulatory capacity, as well as a strong capacity to prevent water crystallization 55 proline: sperm samples, oocytes or in human endothelial cells 58 (sRBCs); 56 GLC-82, LTEP-a-2, 3T3 and smooth muscle cells; 55 Jurkat cells; 26 and human spermatozoa 59 proline + trealose: preservation of cell morphology 56 glutamine: spermatozoa, including humans, 60 boar, 61 ram 62 and rooster, 63 and epididymal sperm in Egyptian spiny mice 20 glutamine: improves motility and protects membrane integrity against damage caused by the freeze−thaw process 20 neutral amino acid-based (β-alanine, γ-aminobutyric acid (GABA), and ε-aminocaproic acid (ε-Ahx)) CPA solutions: cryopreserve GLC-82 cells and SMCs 64 betaine: hydrophilic molecule that has a high capacity for hydration through the ionic solvation effect with its charged groups (inhibitor of ice formation) intracellular ice. However, if the dehydration is very high, then irreversible damage to the cell can occur because of cryopreservation.…”

Dimethyl Sulfoxide: An Ideal Electrochemical Probe for Hydroxyl Radical Detection

“…As a so-called universal solvent, dimethyl sulfoxide (DMSO) is a widely used organic solvent in the fields of chemistry, pharmacology, and biology. In cell biology, DMSO is often used as the cell cryopreservation medium due its ability to restrict ice nucleation and promote post-thaw viability . The DMSO concentration should be reduced to less than 0.1% immediately after cell recovery to prevent its toxic effects.…”