The History of Cryopreservation

Cryopreservation: From Home-Brew to Cutting-Edge Solutions

In the pursuit of medical breakthroughs, technology often emerges from the drive to fulfill critical needs. This is particularly evident in the field of cryopreservation, where a convergence of technological advances seeks to preserve therapeutic cells for a myriad of applications, from treating human afflictions to aiding animals in need. The ability to freeze and revive living organisms has captured the imagination of scientists, philosophers, and dreamers alike. Let’s embark on a journey through the annals of time to explore the evolution, milestones, challenges, and innovations that have shaped the landscape of cryopreservation.

The Fragility of Life and the Birth of Cryopreservation

The roots of cryopreservation trace back to ancient civilizations where tales of suspended animation and attempts to preserve life were woven into mythologies. However, the practical application of preserving biological material through freezing began to take shape in the early 20th century. Cells, once removed from the protective environment of a living organism, become incredibly fragile. Yet, they hold immense potential for repairing host tissues if they can be kept viable. The necessity for prolonged cell preservation outside the body fueled the evolution of cryopreservation science and technology.

Accidental Discoveries: The 1948 Breakthrough

In 1948, scientists Polge, Parks, and Smith stumbled upon a groundbreaking revelation – fowl spermatozoa could be frozen using a concoction of glycerol, albumin, and water.¹ This accidental discovery laid the foundation for cryopreservation, showcasing the potential of home-brewed solutions.

The team aimed to explore the effectiveness of a levulose solution in cryopreserving spermatozoa. An analytical chemist conducted a meticulous analysis to identify the accurate composition of the solution, and further experiments were conducted to confirm that glycerol served as the active ingredient.²

Glycerol and Beyond: The 1950s to 1960s

By 1950, Smith published the use of glycerol for cryopreserving human red blood cells. Lovelock’s work in 1954 evaluated various candidate agents, leading to the recognition of dimethyl sulfoxide (DMSO) in 1959. DMSO became the gold standard cryoprotectant, revolutionizing cryopreservation for various cell types.

1990s – Present: Facing the Limitations

Despite remarkable progress, cryopreservation faces challenges. The formation of ice crystals during freezing can damage cells, and the revival process remains complex. While home-brewed solutions, often comprised of cell culture medium and DMSO, are convenient for low-tech applications, they may present additional challenges in clinical and commercial contexts.

Disadvantages of home-brew:

• Suboptimal cryopreservation: home-brewed solutions may not optimize cryopreservation, leading to unpredictable cell viability and functionality.
• Lack of cGMP compliance: these solutions are not manufactured according to Current Good Manufacturing Practice (cGMP) standards.
• Variable quality: home-brew introduces variability, impacting the consistency and quality of the final cell product.
• Immunogenicity concerns: the use of serum in home-brewed solutions may trigger immune responses in cell therapy recipients.

21^st Century: Cryopreservation in the Modern Age

Advancements in cryoprotectants, nanotechnology, and stem cell research have propelled cryopreservation to the next level. Cryobiologists look to emerging technologies for potential breakthroughs to optimize survival.

CryoStor^®: a leap forward in cryopreservation

BioLife Solutions has translated recent scientific advances into CryoStor, a commercially available cryopreservation solution. This cutting-edge product offers numerous advantages over home-brewed alternatives, including:

• Optimized formulation: CryoStor is an intracellular-like solution designed to minimize apoptosis, reduce ischemia/reperfusion injury, and maximize post-thaw cell recovery.
• cGMP manufacturing: manufactured under cGMP standards using high-grade raw materials, CryoStor ensures consistent quality and performance.
• Off-the-shelf convenience: pre-formulated with 2%, 5%, or 10% DMSO, CryoStor allows users to titrate cell concentrations for optimal survival and function.
• Serum-free and non-immunogenic: CryoStor eliminates immunogenic concerns associated with serum-containing home-brewed solutions.
• Expert guidance: BioLife offers expertise in cryopreservation best practices, assisting in process optimization, freezing protocols, and regulatory compliance.

A future defined by cutting-edge cryopreservation

As we navigate the intricate science of cell cryopreservation, BioLife’s offerings stand at the forefront of innovation. From solution components and freeze-thaw protocols to regulatory expertise, BioLife ensures that every cell in a therapy reaches its full potential. While home-brewed solutions fall short, BioLife’s commitment to quality and innovation provides a reliable bridge from conceptualization to commercialization for cellular therapies. Trust in the advancements of cryopreservation and secure the promise of every viable, functional cell for the future of medicine.

References:

1. Polge C, Smith AU, Parkes AS. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature. 1949;164:666.
2. Pegg DE. The history and principles of cryopreservation. Seminars in Reproductive Medicine. 2002 Feb; 20(1):5-13
3. Smith AU. Prevention of haemolysis during freezing and thawing of red blood cells. The Lancet. 1950;2:910–911.
4. E. LOVELOCK. The Protective Action of Neutral Solutes Against Haemolysis by Freezing and Thawing. The Biochemical Journal. 1954; 56(2): 265-270
5. E. LOVELOCK & M. W. H. BISHOP. Prevention of Freezing Damage to Living Cells by Dimethyl Sulphoxide. Nature. 1959; 183: 1394-1395.
6. Mary C. Phelan. Unit 1.1 Basic Techniques for Mammalian Cell Tissue Culture: Freezing Human Cells Grown in Monolayer Cultures. Current Protocols in Cell Biology (1998) 1.1.1-1.1.10. Copyright © 1998 by John Wiley & Sons, Inc.
7. (2018). Biopreservation Today. Retrieved January 4, 2024, from https://www.biolifesolutions.com/wp-content/uploads/2022/11/BPT-Spring-2018-WEB.pdf.
8. Baust JG, Gao D, Baust JM. Cryopreservation: An emerging paradigm change. Organogenesis. 2009 Jul;5(3):90-6. doi: 10.4161/org.5.3.10021. PMID: 20046670; PMCID: PMC2781087.