Cryopreservation is a vital component of cell therapy and biobanking. In many workflows, the volumes are small, often just tens of milliliters. Whether splitting a unit into aliquots for multiple doses, running early-stage clinical trials with limited cell yields, or banking precious research samples, small-volume cryopreservation presents unique challenges and needs. In this blog post, we explore why specialized freezing bags like the OriGen Biomedical^® CryoStore Freezing Bag (in the 10–30 mL range) are essential for these applications, and how the CryoStore Freezing Bag CS50 stands out as an ideal solution for small-volume storage. We’ll also highlight key technical features, safe handling practices to prevent bag breakage, and accessories that ensure a closed, secure process from freezer to thaw.

Why Small-Volume Cryopreservation Matters

Many cell therapy protocols and laboratory workflows require freezing relatively small volumes of cell suspensions. A single therapy dose or aliquot may be only 20 mL, or a collected unit of cells may be divided into smaller 20-25 mL portions for testing or storage. Using a large cryogenic container for such a small volume can be inefficient and may increase the risk of variability, such as wasted space or uneven freezing. Small-volume freezing bags are designed to address this by providing the right capacity for these use cases, allowing cells to be distributed more evenly during freezing.

From a logistical standpoint,  10–30 mL cryopreservation bags simplify handling by fitting into standard freezing cassettes and inventory systems without the bulk of larger-volume containers. This makes them especially useful for pediatric therapies, decentralized trial sites working with smaller batch sizes, and any setting where dose-splitting or aliquoting is performed. In short, small-volume cryostorage tools help laboratories maximize efficiency and maintain product integrity when managing low-volume but high-value cell products.

The OriGen Biomedical CryoStore 50 Freezing Bags: Designed for Small-Volume Freezing

The CryoStore line includes a small-volume option: CS50, with six total configurations with various tubing options and bag quantities. These bags are engineered specifically for freeze volumes in the 10–30 mL range, making them well-suited for dose-splitting, pediatric applications, and any workflow where only small amounts of cellular material are available.

The CryoStore 50 Freezing Bag uses the same durable EVA film that has been validated for cryogenic use down to −196°C. It incorporates hermetically sealed spike ports for secure access, a corner-positioned donor tube connection to make de-bubbling and sealing easier, and multiple tubing set choices. Every CS50 bag’s donor tubes include pre-printed 100 µL segments to support precise aliquot tracking.

The CryoStore Freezing Bag CS50 and all CS50 configurations are FDA 510(k) cleared, CE Marked, ISO 13485 certified, and manufactured under GMP guidelines. Each bag is provided sterile with a Certificate of Conformance, offering users confidence in consistent performance and regulatory compliance.

Engineered for Ultra-Low Temperature Durability

OriGen Biomedical^® CryoStore Freezing Bags, including the CS50, are validated for cryogenic storage down to −196 °C. The EVA film used in these bags maintains flexibility at ultra-low temperatures, but like all polymers, it becomes more brittle as it approaches its glass transition point. In this state, even durable EVA can fracture if subjected to sudden impact or excessive stress.

To support product safety during storage, CryoStore Freezing Bags can be paired with the OriGen Biomedical O-Wrap^® Bag for an additional layer of protection. CryoStore bags have been validated for short-term resistance to 100% DMSO. When used within recommended concentrations and validated protocols, bags maintain integrity when frozen to cryopreservation temperatures and thawed to 37°C.

Best Practices to Prevent Breakage and Ensure Consistency

While the material is engineered for cryogenic use, proper handling is critical once the bags are frozen. OriGen Biomedical highlights these practices in our white paper How to Prevent Freezing Bag Breaks:

• Seal donor tubes short to reduce leverage stress at the junction.
• Remove air bubbles before freezing, since trapped nitrogen can rapidly expand during thawing and create internal pressure.
• Use the correct freezing cassette so the bag fits securely. Oversized cassettes allow rattling, while undersized ones can pinch the film.
• Handle frozen bags like fragile glass, avoiding bending, dropping, or twisting once below −60°C.
• Follow validated cryoprotectant protocols, as certain agents and trapped gases can create internal stress if not properly managed.
  

The full white paper is available through our Client Dashboard, your hub for technical documents and best practices. For a closer look at the dashboard, its benefits, features, and tips for navigating this exclusive customer portal, visit our blog post Maximizing Your Access to Resources: A Guide to the Client Dashboard.

Applying these precautions helps support consistent performance and the long-term integrity of frozen samples during LN₂ storage.

Accessories for Closed-System Cryopreservation

OriGen Biomedical provides a full accessory line to support closed-system workflows in cryopreservation:

• Cell Connect^® Accessory: Designed for closed-system fluid transfer of cells and cryoprotectant solutions into multiple CryoStore Freezing Bags, using either sterile docking or luer/spike connections.
• O-Wrap^® Bags: Transparent secondary overwraps that serve as an outer container during LN₂ storage and contain leakage in the event of a primary bag failure, helping reduce risks such as thaw bath contamination.
• Tube Sealer CR6: A portable sealer can seal PVC, PVC/EVA coextruded tubing, and EVA tubing, creating consistent, sterile-ready seals that allow secure sample segmentation and traceability.
  

These accessories are CE Marked, ISO 13485 certified, and manufactured under GMP guidelines, ensuring compatibility with OriGen Biomedical freezing bags.

Conclusion

Small-volume cryopreservation relies on precision, consistency, and the right tools. The OriGen Biomedical CryoStore Freezing Bag CS50 brings together durable EVA construction, manufacturing under globally recognized quality standards, and workflow-oriented design features. Paired with validated handling practices and OriGen Biomedical accessory sets, these bags are designed to support the cryopreservation of small cell volumes.