Navigating the Challenges of Cryopreservation in Cell-Based Research
Biopreservation, and specifically cryopreservation, is a critical process for maintaining the viability of cells long-term throughout storage and research. Recently, BioLife Solutions partnered with Cell and Gene Therapy Insights to produce a poster that highlights many of the challenges researchers face, balanced with Biopreservation Best Practice suggestions.
As many know, building and optimizing a cryopreservation protocol to maximize cell viability can be a bit of an art form. While there is no standard method that works for all cell types, we believe the best practices summarized in the poster and outlined below, are a good place to start. For further support and detailed protocol assistance, visit our Ask The Scientists page to be connected directly with our BioLife Bench Scientists.
The Importance of Cryopreservation
Cryopreservation involves cooling cells from biological materials to sub-zero temperatures to halt all biological activity and preserve the cells for future use. These biological materials often originate from tissue biopsies, blood and bone marrow. The materials must remain viable and functional throughout manipulation, manufacturing, storage, transport and administration, to best help patients fighting cancers, auto-immune diseases and genetic disorders.
Biopreservation Best Practices
- Sample Isolation and Cell Processing
Retrieve donor samples at the clinical site and ensure they are packaged and shipped to the manufacturer within scheduled processing times and set development parameters. ** Adding cryoprotectants like HypoThermosol® FRS or CryoStor® CS10 directly to the apheresis material can extend sample shelf life, allowing more time for the sample to reach the manufacturing facility. **
- Formulation
Perform purification and concentration steps to prepare the final drug product. The reduced cell culture should be suspended or formulated in cryopreservation media to ensure optimal preservation.
- Aliquot and Fill
Use commercially available fluid transfer systems, like the Signata CT-5™, to aliquot and fill primary containers, like the CellSeal® CryoCase™. Both automated and manual methods are viable, but maintaining solution agitation throughout this step is crucial to ensure homogeneity and to avoid issues such as sinking cells or coagulation.
**Choosing the right primary containers is critical. Ensure that your selections meet performance, quality, and regulatory requirements. Containers should be reviewed for integrity, extractable and leachable profiles, and scalability for downstream processes.**
- Controlled-Rate Freezing
Utilize a controlled-rate freeze to introduce a “seeding” or ice-nucleation step, which reduces the risk of a supercooling event, and improves sample consistency and process reproducibility. **It’s essential to optimize and qualify the cooling rate according to cell type, but –1°C per minute is a common starting point.**
- Storage and Shipping
Define and qualify storage conditions, including temperature and humidity, to maintain sample integrity and viability. **Partner with biostorage services, like SciSafe Inc., to manage your growing sample storage needs and utilize secure, monitored shipping solutions.**
Secure transport of biological material demands solutions with uncompromised thermal integrity and real-time payload visibility. **Use a shipper like the evo® DV10 for up to 15 days of cryogenic protection, ensuring the sample’s safe arrival.**
- Thawing
Develop a consistent thawing program that can be reproducible at any clinical site. Automated mechanical thawing systems, like the ThawSTAR® CFT2, are preferred over water baths or bead baths to limit site variability and maintain control over the thawing process.
Cryopreservation is a complex but essential process in the development of cell-based therapies. By adhering to best practices and addressing specific challenges, it’s possible to maintain cell viability and functionality, ensuring that these innovative therapies can reach patients safely and effectively. As the field of cell and gene therapy continues to evolve, so too must our approaches to cryopreservation, always striving for greater efficiency and reliability in preserving the very cells that hold the potential to revolutionize medical treatments.
If you are looking for additional resources on biopreservation best practices, review these links:
- https://evidence.biolifesolutions.com/resource/process-optimization-for-cd3-t-cell-formulation-and-cryopreservation
- https://evidence.biolifesolutions.com/resource/delineating-the-critical-process-parameters-for-cell-therapy-cryopreservation
- https://www.insights.bio/cell-and-gene-therapy-insights/journal/article/410/Biopreservation-Best-Practices-for-regenerative-medicine-GMP-manufacturing-focus-on-optimized-biopreservation-media
- https://www.insights.bio/cell-and-gene-therapy-insights/journal/article/1805/Biopreservation-and-cold-chain-biologistics-risk-points-in-the-cell-and-gene-therapy-workflow
