The rise in biomedicine has paved the way for many medical methods to evolve. Cryopreservation, for instance, is now widely used in biological medicine. Cryopreservation involves organ transplant, regenerative medicine, and the discovery of new drugs. And all of these possibilities are only the beginning, the future of cryopreservation seems to be more promising for medical science.
The medical problem of preserving donated organs has catapulted cryopreservation as a technique to address it. There are many applications in cryopreservation that tackles the main challenges in clinical application:
- scaling up to large volumes and complicated tissues
- prevention of ice formation
- alleviating cryoprotectant toxicity
In general, cryopreservation pertains to storing biological materials at low temperatures. Preservation of biological materials for long periods at cold temperatures is present in history. Cryopreservation is now an established science of keeping natural components. Preserving the quality of these is beneficial in medical applications.
Medical Applications
The future of cryopreservation has an extensive field of application. This science is particularly significant in biomedicine.
Organ Preservation
In organ transplants, other than compatibility, the quality of organs is of the highest priority. But because of a lack of preservation capacities, organ transplant procedures in limited. Though, we can possibly see more advanced preservation methods brought by th future of cryopreservation to be the concrete solution.
With better preservation methodology, the effectiveness of organ transplants becomes higher. This will benefit thousands of patients waiting for a suitable organ donor. Better organ preservation facilities cut the queue and streamline transplant operations.
Pharma Research
One organ component that is needed in drug discovery is tissue slices. The tissue samples serve as a proxy as they are a good representation of the organ being tested. Through the use of tissues and cells, research on biological processes can proceed. The future of cryopreservation will serve with high interest in the pharma industry. The researches can gain significant benefits such as:
- warrant big-scale drug screening on human tissues
- identify adverse effects of drugs
- revolutionize toxicology screening
- pave the way for advanced drug discovery
Methods of Cryopreservation
Freezing
The freezing method typically applies to a wide range of the sample. But it is important to note that freezing should follow a consistent colling rate. An ultra-low cooling rate can lead to damage to tissues and cells. Freezing also needs to follow a slow cooling process. Slow cooling gives ample time for water to exit the cells. Such a method will negate the formation of intracellular ice in the tissue.
Cooling standards range to about 1°C per minute. But do note that different cell samples have different optimum cooling conditions.
Vitrification
Vitrification is done through an ultra-low temperature condition. The temperature ranges from -80 to -130 0C. On the contrary, vitrification is a different physical process occurring in the glass transition temperature. In this process, samples can solidify but negate the formation of any ice crystals.
The Cryogenic Process
Cooling
The first process sees the freezing of extra water in the cell tissues. The cell itself loses water due to osmosis. This process pertains to the slow cooling activity. For fast cooling, the intracellular waters will freeze. Freezing occurs because the relay of water in between membranes is not fast enough.
Warming
Cooling the sample first passes through an “ice growth” phase. During a warming state, ice crystal formation is seen around the initial nucleus. This is called nucleation. Such occurrence is dependent on temperature conditions. When the sample passes through non-optimum temperature for crystallization, it enters the nucleation zone.
Cryopreservation Important Ingredients
Nucleation Temperature
There are different levels of nucleation. Primary nucleation happens before the initial formation of ice crystals in the sample. The temperature during primary nucleation is referred to as the nucleation temperature.
Why is it significant? Nucleation temperature affects cell survival. Any extreme temperature condition can be detrimental to a cell. There are many ways to control the process of nucleation.
- application of cooled metal rod
- expanded gases
- application of ice-nucleating agents
In the case of ice-nucleating agents, the application is minimal. Any extra amount can lead to toxicity.
Cryoprotective Agents (CPAs)
CPAs are significant in cryoinjury prevention. The use of CPAs can reduce solution effects. Using a good CPA is tantamount to standardizing cryopreservation as a whole. The properties of good CPA include:
- water-soluble at low temperature and high concentrations
- can properly penetrate cell membranes
- low toxicity
Toxicity is a widely known challenge in cryopreservation. The future of cryopreservation would look into negating such toxicity levels of CPAs. This is because the toxic effects of CPAs are affected by temperature.
Biological Anti-Freeze Proteins
Antifreeze proteins are present in animals, especially those living in cold regions. The substance allows these animals to survive the cold temperature. Antifreeze proteins can bind to ice and prevent it from growing.
Synthetic Ice Blockers
Because antifreeze proteins are expensive, there are synthetic options available. Synthetic ice blockers directly bind to ice. The use of these ice blockers limits the heterogeneous ice nucleation in the process.
Compatible Solutes
They are also called ice blockers. They stop the formation of ice when used. Compatible solutes are derived from a microorganism. This microorganism produces solutes under stress.
Vitrification Solution
These solutions allow larger samples to be vitrified. In the simplest form, it turns pieces into a glassy substance. A more concentrated vitrification solution is needed to preserve larger tissues. A more significant concentration is required to cover a bigger surface area ratio.
It is important to note that high concentrations of solutions can be toxic. This is why specific measures are made to reduce it. Often, cryoprotectants are mixed in a solution to reduce the toxicity level. By combining different CPAs, it reduces the concentration of a single CPA present. That single CPA may be the cause of toxicity.
However, it is one of the hopes that this won’t be needed in the future of cryopreservation.
Cryopreservation Equipment
Freezer
An ultra-low temperature freezer can help in maintaining controlled rate cooling. If the need is for repaid freezing, a blast freezer is a suitable option. Also, a different kind of freezer is best for specific samples. Blast freezers are best for plasma storage and bioproduction. A generic laboratory freezer can store robust cells such as lab stocks, proteins, and other non-critical samples. On the other hand, a controlled rate freezer is suitable for stem cell operations. It is also best for storing embryo, IVF, and DNA samples.
Storage
The general principle in cryopreservation is the lower the storage temperature, the longer the viable storage period will be. Storage has precise guidelines also depending on the samples being preserved. Intracellular components need storage at -70 0C. This can last for months and years. Within this temperature range, chemical reactions are not entirely stopped. But samples stored at -130 0C are preserved indefinitely.
The Future of Cryopreservation
There are extensive possibilities for the use of cryopreservation. In clinical applications alone, cryopreservation serves a significant impact. Cryopreservation of blood components is one concrete use case.
With rigid prerequisites in cryopreservation, particular innovation in equipment is following suit. The advent of innovative freezer technologies has made rise to better preservation outcomes. While these developments are promising, specific preventive measures are a relevant topic.
To streamline the scientific methodology, monitoring is imperative. Monitoring accounts for the optimum condition of the equipment. This enables the maintenance of consistent temperature conditions specific to biological samples.
AKCP Monitoring Systems
Wireless Temperature Monitoring
Temperature monitoring for cold storage equipment can be done in real-time. Wireless temperature sensors from AKCP log data and graphs regularly. The data are programmed with pre-defined thresholds that will raise alerts when critical conditions arise.
A cryo facility monitored with wireless sensors enables granular visibility capable of remote checking. A bonus is when sensors are connected to a monitoring software for central tracking. Live central monitoring of the deployed devices in a cryo facility is a robust monitoring solution that assures the quality of sample preservation.
AKCPro Server is an interface that tracks all placed sensors in the blast freezer, for example. It generates daily reports and graphs as evidential documents for easy reference.
PT100 Type Ultra Cold Temperature Sensor
This sensor is suitable for monitoring a wider range of temperatures from -200°C to +150°C (-328°F to 302°F). Used for ultra-cold storage, such as the mRNA based COVID-19 vaccines from Pfizer and Moderna. The Ultra Cold Temperature Sensor is based is an Resistance Temperature Detector type sensor made from Platinum. As temperature changes so does the resistance of the metal. This is precisely measured to give a temperature reading. The accuracy of the sensor is ±0.2°C (±0.4°F).
- 4x AA Battery powered, with 10-year life.
- USB 5VDC external power.
- 12VDC external power.
- Custom sensor cable length up to 15FT. to position sensor in optimal location.
Wireless Tank Monitoring
Cloud-based monitoring system makes use of telemetry technology and sensing devices to offer cryogenic level monitoring solution. Wireless Tank Level Sensor simplifies liquid gas consumption tracking, processing, and real-time monitoring. It enables the gas companies to inspect the present status of cryogenic containers as its availability and delivery is crucial and time-sensitive.
Wireless tank monitoring system resolves these issues through automatic monitoring that reduces human intervention and allows potential cost savings and valuable time. Not only the tank level is accurately tracked but automatically updated to the central system, eliminating paperwork.
The cryogenic storage poses many challenges to healthcare facilities. But with the help of AKCP monitoring maintaining protocol compliance is made easy through precision monitoring and thorough documentation. With the environmental monitoring solutions and proactive alerts within the administrators.
Realize the benefits and reliability of digital technology for storing specimens, ultra-low freezing, manufacturing in addition to real-time monitoring, alarming, and automated reporting.
Monitor Fuel Level and Liquid Storage Tanks
Monitor tanks of depths up to 20 meters. Often tanks are located in outdoor or difficult to cable areas. The WT-TDPS is battery powered, or can be powered from a 5VDC or 12VDC source. Track usage, graph the tank level, receive alerts when tank levels are critical. No more constraints on maximum cable lengths from the base unit. Easy installation and pairing with AKCP Wireless Tunnel Gateways.
- 4xAA Battery powered, with 10-year life.
- Tank mounting kit with cable gland
- IP66 rated enclosure
- Monitor all kinds of liquids.
Clinical Breakthrough
Organ transplant has made enormous strides in advanced health methods. Even reproductive health methods such as IVF and egg cell freezing are dependent on cryobiology. These advancing demands are a clinical breakthrough that will push further in the future.
The need to scale up to preserve larger volumes and more complex tissues is closer to fruition. That said, a combination of equipment innovation and robust monitoring solutions is required. The small wins in the cryobiology field are thanks to dedicated research. From mitigating cryoprotectant toxicity to limiting ice formation, tiny details are of proportionate significance to the bigger picture.
There is a future of opportunities. We are in for a ride of breakthroughs after breakthroughs.
Reference Links:
https://bmcbiol.biomedcentral.com/articles/10.1186/s12915-021-00976-8
https://www.open.edu/openlearn/body-mind/health/health-studies/what-biomedicine
https://www.technologynetworks.com/cell-science/articles/cryopreservation-freezing-methods-and-equipment-292855
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