Cryopreservation is the use of low temperatures to preserve structurally intact living cells. Cells are cryopreserved to avoid loss by contamination, to minimize genetic change in continuous lines and to avoid transformation in finite lines. Successful cryopreservation of cells depends on optimal freezing conditions, storage, and proper cell thawing techniques. A standardized and reproducible protocol must be followed, although each protocol may require optimization for a given cell type or line, to achieve maximum viability upon thaw. Mammalian cells that are cryopreserved include immortalized cell lines, primary cells isolated from tissues and stem cells. Mammalian cells are best frozen in the presence of a cryopreservant such as DMSO or glycerol, with a freeze rate of 1°C/minute to avoid detrimental ice crystal formation as water within the cell is frozen. BioCision solutions for for cell cryopreservation include CoolBox® XT ice-free cell preparation workstations for temperature controlled sample preparation, CoolCell® alcohol-free cell freezing containers for reproducible controlled-rate freezing, TruCool® cryogenic vials and hinged cryoboxes for sample storage.
Cryogenic vial preparation is typically performed under aseptic conditions in a cell culture hood. Cell suspensions are typically kept cold (0.5 to 4.0℃) which requires the use of ice outside the hood or an ice-free cooling unit inside the hood. The CoolBox® XT ice-free cooling workstations can be used inside a sterile hood environment and will keep up to 48 cryogenic vials cold (0.5 to 4℃) for >16 hours. The thermo-conductive properties of the cooling core inside the CoolBox XT and the cryogenic vial CoolRack holder ensure uniform temperature distribution to all vials during the process. CoolBox XT does not require ice, electricity or batteries.
Optimal cell cryopreservation requires a controlled freezing rate of -1℃/minute for most types of cells and choosing the right freezing method is crucial. CoolCell® alcohol-free cell freezing containers provide controlled rate freezing in a -80℃ freezer. Current methods that utilize isopropanol-based freezing containers and styrofoam boxes do not provide uniform freezing rates to all vials and/or may not be reproducible. CoolCell® alcohol-free cell freezing containers provide a cost effective means of reproducibly conducting the cell freezing process in a -80℃ freezer or with a portable dry ice temperature stability system. CoolCell freezing containers provide uniform, consistent and reproducible cell cryopreservation.
How do CoolCell containers work?
CoolCell freezing containers are passive devices that provide a controlled -1℃/minute freeze rate to all cryogenic vials when placed in a -80℃ freezer. CoolCell containers do not require isopropanol or any fluids - the design and materials regulate heat removal and provide uniform and reproducible freezing to all vials.
Freezing consistency. Five consecutive freeze runs in a -80℃ freezer show highly reproducible freezing profiles.
How do CoolCell containers compare to other freezing methods?
• Consistent -1˚C/minute freeze rate to all vials
• Alcohol-free; no ongoing cost, maintenance or waste
• Reproducible freezing profiles
• 5-10 minute wait period between freeze runs (allows 2 freeze runs per day)
• Optimal for pluripotent stem cell recovery (see above)
Unstandardized; difficult to document rate of cell freezing as Styrofoam containers can vary greatly in size, geometry, density and structure; not reproducible
Stated freeze rate of -1°C/minute, but varies based on vial position; continuous isopropanol replenishment, cost and waste; isopropanol is a variable in each freeze run hindering consistent reproducibility; long wait periods between freeze runs
The device is elegant in its simplicity and ease of use and offers researchers a method to cryopreserve cells in a standardized fashion with great reproducibility and little variability in performance.
- Kevin Grady, ATCC
We run a registry, in which large amounts of PBMCs are processed for long-term cyropreservation. After testing the CoolCell out, we found slightly better cell viability (>90%) than our current cell freezing containers, and there is no Isopropanol waste generated. Overall, the CoolCell has proven to us to set a new bar in cryopreservation.
- Rohit Gupta, Stanford University
The CoolCell is more efficient and easier to use than the Mr. Frosty. Not needing to add isopropyl, the lack of a screw top, and not having downtime after removing it from the -80° makes freezing cells a lot easier. I plan on only purchasing the CoolCell in the future.
- Matt Donne, UCSF Stem Cell Lab
Purely altruistically, I wanted to mention that one big advantage of the CoolCell is that you can use dry ice to do your freezing if you don't have a -80. I spent nearly a year getting terrible viability-recovery on cell freezing with dry ice (in an alcohol device) followed by storage in LN2, and this on HeLa cells which aren't exactly hard to store. I then bought a CoolCell and now I can just use standard cryo-preservation techniques (DMSO, 20% NBS) and get nearly 100% viability on thawing.
- Sam Knight, Ceramisphere (Australia)
 Shu Z, Kang X, Chen H, Zhou X, Purtteman J, Yadock D, Heimfeld S, and Gao DDevelopment of a Reliable, Low-cost, Controlled Cooling Rate Instrument for the Cryopreservation of Hematopoietic Stem Cells. Cytotherapy. 2010.