What is cryopreservation?

Sometimes, We need to cool the samples at ultra-low temperatures to preserves organelles, cells, tissues, or any other biological constructs. When we thaw it. It can be restored to the same viable state as before it was frozen. This process is named "Cryopreservation".

Why use cryopreservation?

The cryopreservation techniques are usually used in biochemistry and molecular biology, food sciences, ecology and plant physiology, reproductive medicine, and many medical applications. The artificial insemination is one of the famous applications.

The cryoprotective agents used in cryopreservation techniques

During the freezing and the thawing cycle, If freezing the cells with simple and cursory cooling, it might lead the cells to be cryoinjury or death. It brings about the membrane damage through the ice crystal formation. Accidentally, In 1948, It was discovered that the ability of glycerol to protect cells from freezing injury. The forerunner of cryoprotective agents was born. In recent years, the use of cryoprotective agents, Increase the survival rate of cryopreservation cells and tissues successfully.

Cryopreservation Procedure can be classified into 4 basic categories.

According to different cell types, the cryopreservation processes can roughly be grouped into fourSlow freezing types:

1)Slow freezing

• The most appropriate storage method of small volumes cryopreservation of several microencapsulated cells.
• It must be carefully selected that the CPA solution, sample preparation, cooling, thawing and CPA removal.

2)Vitrification

• There are three primary constituent in the cryoprotectant. involve Polyvinylpyffolidone (PVP), Dimethyl Sulfoxide (DMSO), and Glycerol.
• It must be used the cryoprotectant concentrations which sufficient to prevent the crystallization of ice altogether.
• High concentrations of cryoprotectants are possibly harmful to cells.
• The lowered concentration can be realized through using very rapid cooling, and even more rapid warming.
• Add the HEPES into the cryopreservative to buffering the pH of the cryopreservative, It also has the effect of making the cryopreservative an isotonic solution of mitochondria.

3)Subzero nonfreezing storage

• It uses a refrigerating device that generates a variable magnetic field.
• Even if without the use of cryoprotectants, the sample would be cooled to a supercooled state.
• It was verified the method has the ability to suppressing anaerobic metabolism and decrease harm in the hearts of large animals.

4)Preservation in the dry state.

• The ultimate goal is fully realized when stable long-term storage without the use of liquid nitrogen and bulky freezers, It becomes available.
• It is prove that increase cell survival at low water contents by loading cells with the protective disaccharide trehalose.
• It needs a low level of residual water is required to achieve viability after rehydration.

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