- A cryoprotective agent (CPA) is a substance used to protect biological materials, such as cells, tissues, embryos, and organs, from damage caused by ice crystal formation during freezing and thawing.
- These agents are essential in cryopreservation, a process that involves storing biological samples at ultra-low temperatures (-150°C to -196°C).
- Without CPAs, ice crystals can form within cells, leading to mechanical damage, membrane rupture, and loss of cellular function.
- CPAs work by reducing the freezing point, minimizing osmotic shock, and preventing intracellular ice formation, thereby improving post-thaw viability. They stabilize cell membranes and maintain osmotic balance, ensuring cell survival during the freeze-thaw cycle.
- CPAs are classified into permeating and non-permeating agents.
- Permeating CPAs like dimethyl sulfoxide (DMSO), glycerol, ethylene glycol, and propylene glycol penetrate cells, replacing intracellular water and preventing ice crystal formation.
- Non-permeating CPAs such as sucrose, trehalose, polyethylene glycol (PEG), and serum albumin remain in the extracellular space, drawing out water to reduce ice formation.
- Different cell types require specific CPAs—DMSO is commonly used for mammalian cells and stem cells, glycerol for bacteria and red blood cells, and ethylene glycol for embryos and oocytes.
- While CPAs are critical for cryopreservation, some, like DMSO, can be toxic at high concentrations, necessitating careful handling and post-thaw removal.
- Advances in serum-free and xeno-free cryoprotectant formulations are improving cryopreservation efficiency for regenerative medicine, fertility preservation, and cell therapy.
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