Dimethyl sulfoxide (DMSO) is the most commonly used permeating cryoprotectant in cell cryopreservation, playing an irreplaceable and critical role in the low-temperature storage of cells.
Core Mechanisms of Action:
1. Prevention of ice crystal formation (primary function)
Key principle: Intracellular ice crystals can puncture cell membranes and organelles, leading to cell death. DMSO protects cells by lowering the freezing point and reducing ice crystal formation.
(1) Lowering the freezing point: DMSO rapidly permeates the cell membrane, binds with water molecules, and lowers the freezing point of the intracellular solution.
(2) Inhibiting ice crystals: It reduces ice crystal formation during freezing, preventing sharp crystals from puncturing cell membranes and organelles.
(3) Vitrification: It promotes the formation of a vitreous state (a non-crystalline solid state) in water at low temperatures, thereby mitigating physical damage.
2. Maintenance of osmotic balance
(1) Replacing water: DMSO partially replaces intracellular water, reducing drastic volume changes caused by cellular dehydration during freezing.
(2) Buffering osmotic pressure: It works synergistically with other medium components (such as fetal bovine serum) to buffer osmotic shock during the freeze-thaw process.
(3) Preventing shrinkage/swelling: It prevents cells from shrinking or swelling and bursting due to osmotic imbalance.
3. Protection of cell membranes and protein structures
(1) Stabilizing cell membranes: It interacts with membrane lipids to form a stable lipid bilayer structure, preventing membrane rupture caused by reduced fluidity at low temperatures.
(2) Preventing protein denaturation: It reduces protein denaturation caused by elevated electrolyte concentrations and pH changes during freezing.
(3) Antioxidant effect: It possesses antioxidant properties, scavenging or reducing reactive oxygen species (ROS) generated during the freezing process.
4. Increasing intracellular ion concentration
(1) Increasing viscosity: It increases the viscosity of the intracellular solution, reducing the mobility of water molecules.
(2) Retarding the freezing process: It slows down the freezing process, allowing cells more time to adapt to the low-temperature environment.
(3) Reducing damage: It minimizes intracellular ice crystal formation, thereby reducing cellular damage.
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