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Everything You Need to Know About Cryopreserved Leukopaks

What Is a Cryopreserved Leukopak?

A cryopreserved leukopak is a leukopak that is frozen at a subzero temperature (normally -196 degrees Celsius) to preserve the biological sample for long-term storage. A leukopak is collected via leukapheresis and consists of white blood cells, plasma, platelets, and red blood cells. 

Leukopaks are used for immune studies, cell therapy and have several other biomedical applications. They are particularly excellent experimental resources because they contain more than six times as many leukocytes—white blood cells—and more than 20 times as many human peripheral blood mononuclear cells (PBMCs)—monocytes and lymphocytes – compared to the same volume of whole blood. Researchers will isolate  these leukopaks into their components via blood separation techniques and cell separation techniques.

When conducting these experiments, researchers must wait for daily shipments of fresh cells in the form of leukopaks until they begin a processing protocol. Delays in shipments of fresh cells can result in a race against time, increased degradation of cell quality, squandered research funds, and inaccurate results. 

Cryopreserved leukopaks eliminate the stress of shipments or delayed transit by prolonging the leukopak’s shipping and storage durability. By cryopreserving the leukopak, the amount of time a leukopak can remain useful increases tenfold, guaranteeing a maximum of a 2% viability loss. 

What Is in a Frozen Leukopak?

A cryopreserved leukopak is a hyper-frozen leukopak. Cryopreservation typically happens at -196 degrees Celsius and essentially instantly stops the biochemical processes of the cells within. The quality of the leukopak is preserved in a moment. Similar to fresh leukopaks, cryopreserved leukopaks are varied in contents depending on the supplier, the success and type of apheresis, the intended use, and the donor quality. 

Generally, cryopreserved leukopaks are made up of 50% T cells, 20% monocytes, 10% B cells, 10% natural killer (NK) cells, 3% granulocytes, and 3% hematocrit. Mobilized leukopaks can contain even higher concentrations of these elements. All of these components can be separated from leukopaks for research and are often divided based on the experiment at hand. 

How Long Will a Cryopreserved Leukopak Stay Frozen?

Fresh unfrozen cells’ viability decreases rapidly after collection until about 24 hours post collection when the majority of cells are no longer viable. Shipment delays can have catastrophic effects on the quality of fresh leukopaks.  During shipment on dry ice, a cryopreserved leukopak will remain frozen for up to a month, eliminating the stress associated with shipping live cells long distances.

Once a shipment of cryopreserved leukopaks arrives at a facility, they can be stored for a month in a -80 degrees Celcius freezer and up to a year in liquid nitrogen . Fresh leukopaks have a short shelf life, often dictating when researchers will conduct experiments, which can lead to rushed processes. Researchers can better plan and execute with the added stability of cryopreserved leukopaks. 

Benefits of Cryopreserved Leukopaks 

Ability to Transport

Transporting fresh cells globally is an extremely complex process that too often entails a significant loss in quality of cells and experiments, lab funds, and time. Cryopreserved leukopaks eliminate the inherent risk of long shipping distances or shipping delays with no decrease in quality.

Maintain Sample Quality

It is important to maintain the quality of the leukopak throughout shipping, storage, and experimentation. The cryopreservation process essentially locks the living cells at a point in time, stopping the course of degradation. The health of the cell population at the critical point when the cells are frozen is a key factor in determining the health and viability of the cell population post-thaw. Most cryopreserved leukopak providers will guarantee at least 95-98% cell viability survival, a promising trade-off for increased storage and shipping leniency. 

Easy Storage

Cryopreserved leukopaks keep for up to a month in a -80 degrees Celsius freezer and 1.5 years in liquid nitrogen. Cryopreserved leukopaks are more than 10 times more resilient than fresh cells and can be thawed when needed, conserving the quality. .

Thawing a Cryopreserved Leukopak 

Cryopreserved leukopaks should be thawed in a warm water bath and inverted three or four times until homogenous. In some cases, this can take as little as two minutes. Once the contents have been properly thawed, they can be kept on ice for 24 hours until assayed or conducting a cell separation experiment. 

This flexibility in storage and thawing procedure allows researchers to pull a cryopreserved leukopak from the reagents freezer—along with their other reagents in use—and have a viable cell solution ready to use in moments. 

Akadeum’s Cell Isolation From Commercial Leukopaks

Once the cryopreserved leukopak has thawed, Akadeum products can assist with simplifying the separation steps. Akadeum specializes in making cell separation simple using our specifically engineered naturally buoyant microbubbles. Microbubbles bind to unwanted cells and then they both rise to the top of the leukopak solution. Meaning the margin of error from harsh cell environments, switching between tubes, or pour-offs will no longer be an issue. 

Using Akadeum microbubbles, a time-consuming and risky preparation step can become a hardy protocol with high yield and dependable delivery, allowing the researcher to focus on the science at hand. The process can be applied to whole leukopaks, ensuring the effortless preparation of isolated cell products for research. 

Whether extracting cellular subsets from a buffy coat, a leukopak, or a cryopreserved leukopak, Akadeum’s Microbubble cell separation kits will deliver focused and accurate cell isolation every time. 

If you want to know more about how our microbubbles can provide quick and easy cell isolation, explore our Human T Cell Isolation Kit to see our technology in action. 

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