Cryopreservation in Cell and Gene Therapy: Ensuring High-Viability Cells for Research & Manufacturing

By Joanna Wirkus

Estimated reading time: 6 minutes

The Role of Cryopreservation in Cell and Gene Therapy

Cryopreservation is a cornerstone of cell and gene therapy, enabling the long-term storage and viability of vital cell types such as CD34+ hematopoietic stem cells, mesenchymal stem cells (MSCs), natural killer (NK) cells, and T cells used in CAR-T therapies.

Interestingly, nature has its own version of cryopreservation: the North American wood frog survives winter by producing natural cryoprotectants like glucose and urea-no liquid nitrogen required.

How Cryopreservation Works

Water’s hydrogen bonds cause ice crystals to form at low temperatures, which can puncture cell membranes and kill cells. Cryoprotectants like dimethyl sulfoxide (DMSO) prevent this by:

• Disrupting hydrogen bonding to stop ice crystals from forming
• Lowering the freezing point to reduce intracellular ice
• Stabilizing cell membranes through lipid interactions

The foundation for modern cryopreservation began in 1949 when Christopher Polge accidentally discovered that rooster spermatozoa could survive freezing in glycerol. Today, this process is central to biobanking, clinical-grade manufacturing, and gene-modified therapies.

CGT Global’s Optimized Cryopreservation Process

At CGT Global, we provide high-viability, cryopreserved primary cells for preclinical research, clinical trials, and commercial-scale therapy development. Our process includes:

• Cell-Specific Cryopreservation Media – Tailored formulations with DMSO and supplements, designed for maximum recovery when paired with our optimized thawing protocol.
• Controlled-Rate Freezing – Data-driven cooling protocols that minimize ice crystal damage and osmotic stress.
• 7-Year Stability Guarantee – Cells retain high viability when stored and thawed properly, ensuring reliability over long-term projects.
• Customizable Protocols – Need non-standard cryoprotectants or storage conditions? We’ll tailor a solution for your project.

Cryopreserved Cells Available for CGT Applications

We offer a diverse portfolio of isolated, cryopreserved primary cells critical for cell and gene therapy manufacturing:

• CD34+ Hematopoietic Stem Cells – Key for gene-modified therapies, CAR-T, and transplantation
• Mesenchymal Stem Cells (MSCs) – Essential for regenerative medicine and immune modulation
• T Cells & CAR-T Cells – Core to cancer immunotherapy platforms
• Natural Killer (NK) Cells – A promising option for off-the-shelf cancer therapies

Explore our cryopreserved primary cell inventory for your upcoming research.

Validated, Temperature-Controlled Global Shipping

We make sure your cells arrive viable and ready to go:

✔ Large, consistent lot sizes for reproducibility
✔ Recallable donor pool with customizable characteristics
✔ Temperature-monitored shipping backed by global logistics expertise

📦 Contact CGT Global today to source high-quality cryopreserved cells for your next breakthrough.

Thawing Best Practices: Preserving Cell Viability Post-Cryo

Cryopreservation is only half the equation—how you thaw your cells can significantly impact post-thaw viability, function, and experimental consistency. Whether you’re working with cryopreserved Leukopaks, PBMCs, or other immune cells, following a standardized thawing protocol is critical.

It must be slow enough to prevent harmful intracellular ice formation, yet fast enough to avoid excessive dehydration and cell shrinkage. To further boost survival, cryoprotectants (CPAs) like DMSO are routinely added to freezing media to protect cells throughout the process.

At CGT Global, we provide a detailed Thawing Instructions Sheet with every cryopreserved product to help you minimize variability and maximize recovery.

Key Tips for Successful Thawing:

• Rapid Thawing: Use a 37°C water bath and remove the vial promptly once thawed—don’t let cells sit too long at elevated temperatures.
• Gentle Handling: Avoid harsh pipetting or vortexing; these can damage delicate immune cells.
• DMSO Removal: Dilute and wash cells gradually to reduce DMSO toxicity.
• Recovery Period: Allow cells to rest post-thaw before downstream applications (e.g., overnight incubation).

Need help? Download our Cryopreserved Cell Thawing Protocol

Leukopaks Thawing Protocol

Primary Cells Thawing Protocol

The Future of Cryopreservation: Innovations and AI Advancements 

1. Emerging Cryopreservation Technologies 

Cryopreservation is advancing with cutting-edge innovations designed to improve cell viability and reduce cryoprotectant toxicity: 

• Ice-Free Vitrification – A technique that avoids ice crystal formation altogether by rapidly cooling cells into a glass-like state. 

• Nanoparticle-Based Cryoprotectants – Alternative cryoprotectants designed to reduce DMSO toxicity, especially for clinical applications. 

• Automated Freezing Systems – AI-driven protocols that optimize cooling rates for maximum post-thaw viability. 

2. Overcoming Cryopreservation Challenges in Cell and Gene Therapy 

While cryopreservation is essential, it presents unique challenges: 

• DMSO Toxicity – Can negatively impact sensitive cell types. Some researchers are exploring DMSO-free cryopreservation. 

• Post-Thaw Recovery Rates – Some cells are more vulnerable to cryo-damage. Optimized thawing protocols are crucial. 

• Scalability for Cell Therapy Manufacturing – As therapies move from lab to clinic, maintaining consistent lot quality at scale is a growing concern. 

3. Cryopreservation & Space Biology 

Did you know that NASA and SpaceX are researching cryopreserved stem cells for space travel? In microgravity, stem cells behave differently, and cryopreservation is key to future off-world medicine and biomanufacturing. 

4. AI in Cryopreservation: Optimizing Freezing & Thawing Protocols 

• AI-driven predictive modeling determines the ideal cooling and warming rates for different cell types, reducing ice crystallization damage. 

• Machine learning algorithms analyze post-thaw viability data to improve cryoprotectant formulations. 

• Automated cryopreservation systems use AI to adjust freezing rates in real-time, improving reproducibility. 

5. AI for Predicting Post-Thaw Cell Viability 

• AI models trained on thousands of cryopreserved samples predict post-thaw viability based on donor characteristics, storage conditions, and freezing parameters. 

• This helps biotech and pharma companies reduce waste by selecting the best preservation conditions in advance. 

6. AI in Cryo-Logistics: Smarter Supply Chains for Cell Therapy 

• AI-powered real-time monitoring of cryoshipments detects temperature fluctuations and predicts risks to cell viability. 

• AI improves inventory management for biobanks and cell therapy manufacturing, ensuring high-demand cells are always available. 

Ready to harness the future of cryopreservation?
Let’s build smarter, more scalable cell therapy workflows-partner with CGT Global today.

Sources

Polge, C., Smith, A., & Parkes, A. (1949). Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 164(4172), 666. https://doi.org/10.1038/164666a0 

Food and Agriculture Organization of the United Nations (FAO). (2012). Basic principles of cryopreservation. https://openknowledge.fao.org/server/api/core/bitstreams/4b22ef40-3af1-4252-a817-92b534dc0116/content/i3017e04.pdf 

NASA. (2020). BioScience-4: SpaceX CRS-16 & SpaceX CRS-21 research on stem cells in microgravity. https://www.nasa.gov/general/bioscience-4-spacex-16-spacex-21/ 

Higgins, A. Z., Karlsson, J. O. M., & Toner, M. (2021). Nanoparticle-mediated delivery of cryoprotectants for cryopreservation. Frontiers in Bioengineering and Biotechnology, 9, 632677. https://doi.org/10.3389/fbioe.2021.632677 

Baust, J. G., Corwin, W. L., Van Buskirk, R. G., Baust, J. M., & Snyder, K. K. (2019). The impact of varying cooling and thawing rates on the quality of cryopreserved human peripheral blood T cells. Cryobiology, 87, 50-57. https://pubmed.ncbi.nlm.nih.gov/30833714/ 

Costanzo, J. P., Lee, R. E., & Wright, M. F. (2006). Cryoprotection by urea in a terrestrially hibernating frog. Journal of Experimental Biology, 209(3), 467-477. https://pubmed.ncbi.nlm.nih.gov/16244167/ 

Puhlev, I., Guo, N., Brown, D. R., & Levine, F. (2001). Lyophilization as an alternative to cryopreservation. Cryobiology, 43(3), 293-306. https://parentsguidecordblood.org/en/news/lyophilization-alternative-cryopreservation