Cell Cryopreservation: Techniques, Applications and Significance

 

Cell Cryopreservation

What is it?

 Cell cryopreservation refers to the process where cell viability and functionality are preserved by cooling the cells to very low sub-zero temperatures typically using solid carbon dioxide, liquid nitrogen or other cryoprotectants. At such low temperatures, any biological or chemical activity including metabolism, growth, and degeneration that may damage the cells is effectively halted. This enables long-term storage and preservation of cells so that they can be retrieved or 'thawed' when required without loss of quality for future applications.


Techniques of Cell Cryopreservation

There are two main techniques used for Cell cryopreservation- slow freezing and vitrification. Both techniques aim to cool the cells in a controlled manner to avoid the formation of ice crystals that can damage the cells.

Slow Freezing Technique

The slow freezing technique involves cooling the cells slowly at a controlled rate of around 1°C/min to –80°C before transferring them to liquid nitrogen storage at –196°C. Here, the cells are cooled gradually allowing sufficient time for intracellular water to migrate out of the cells and form extracellular ice. This technique requires the use of cryoprotective agents like dimethyl sulfoxide, glycerol or ethylene glycol along with the cells to prevent intracellular ice formation. These cryoprotectants penetrate the cells and protect them during cooling.

Vitrification Technique

The vitrification technique cools the cells at a very rapid rate of typically 10,000°C/min or higher to transform the solution directly into an amorphous glass-like solid or "vitrified" state in which no ice crystals are formed. Here higher concentrations of cryoprotectants like glucose, sucrose etc. as solutes are used along with rapid cooling to vitrify the cell solution before ice crystal formation can occur. This technique eliminates ice crystal damage but requires rapid cooling equipment and higher cryoprotectant concentrations, which may be toxic for cells.

Applications of Cell Cryopreservation

Cell cryopreservation has wide applications in various fields including medicine, biotechnology and agriculture. Some key applications are:

Stem Cell Banking
Cryopreservation plays a vital role in stem cell banking for future clinical usage. It enables long-term storage and easy access to stem cells when needed for treatments like bone marrow transplants. This is crucial for autologous as well as donor stem cells.

Organ and Tissue Transplantation
Cells from donor organs can be cryopreserved to expand the donor pool and overcome limitations of geographic distance and availability. It helps increase success rates for corneal transplants, cardiac valves transplants etc.

Fertility Preservation
Oocyte and embryo cryopreservation helps preserve fertility for women undergoing cancer treatment or genetic diseases. It allows them a future chance at pregnancy through in-vitro fertilization.

Cell line banking
Established cell lines are cryopreserved in cell banks for long-term storage as 'model systems' in cell biology research along with newly developed cell lines. This facilitates wider distribution and shared use.

Cryopreserved semen banking
It is a key method for long-term preservation and storage of human semen samples, important for fertility treatment, forensic analyses and future reproduction when needed.

Embryo Biobanking
Cryobanking of surplus embryos avoids their wastage and allows future selection for transfer based on the health, genetics or preferences of intended parents later.

Significance of Cell Cryopreservation

Cell cryopreservation has greatly supported advances in various areas due to some key significance:

- Enables long-term storage of biological materials and resources for extended durations, even indefinitely when stored in liquid nitrogen.

- Facilitates broad distribution and sharing of living cells, cell lines and biological products globally through easy transportation in frozen state.

- Provides access or 'off-the-shelf availability' of viable cells when instantly required for research, clinical therapies or other applications.

- Helps overcome limitations of geographic and temporal access through cryopreservation of stem cells, organs, cell lines etc. in bio banks.

- Supports advances in regenerative medicine through utility of cryopreserved stem cells, tissues for eventual therapeutic usage.

- Maximizes utility of biological specimens and resources through their multiuse, retrieval and analysis over prolonged durations.

- Plays a crucial role through fertility preservation for cancer patients and genetic disorder affected individuals.

Cell cryopreservation offers significant technological andscientific advantages through controlled long-term storage of cells without loss of viability or functionality. It is a key enabler facilitating the many advances and applications of modern biomedical research and therapeutics.

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