CryoDefend-Cell Lines (5 x 10 mL)
R&D Systems, part of Bio-Techne | Catalog # CCM019
Key Product Details
Summary for CryoDefend-Cell Lines (5 x 10 mL)
Media for defined, protein-free cryopreservation of cultured cells.
Key Benefits
- Reduce experimental variability by using fully defined media
- Supports post-thaw cell viability better than conventional freezing media
- Optimized for cryopreservation of cultured cells
- Suitable for all cell types
Why use fully defined, protein-free media to cryopreserve cells?
Cryopreservation can be used to maintain cells at a low passage number and to reduce the risk of contamination by microbes and other cell types. The viability of the recovered cells depends on cell density, the type of cryoprotectant used, the method of cooling, and the amount of the cryoprotectant that remains in the culture after the cells have been thawed.
R&D Systems offers CryoDefend® Cell Lines Media, a defined, protein-free media, which has been optimized for the cryopreservation of cultured cells. This specially formulated media contains a defined serum substitute as well as an optimized concentration of a cryopreservative that increases the recovery and viability of healthy cells compared to conventional freezing media.
Components
- 5 x 10 mL vials of CryoDefend® Cell Lines Media
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Cell Viability Following Cryopreservation in CryoDefend®Cell Lines Media in Comparison to Conventional Cryopreservation Media. A variety of cell lines were frozen in triplicate in either control freezing media (90% culture media/10% DMSO) or CryoDefend®Cell Lines (CryoDefend®-CL) media at 1 x 106cells/cryovial and stored in liquid nitrogen for one week. The cells were then thawed, counted (blue bars), resuspended in standard culture media, and plated into T75 flasks. After 3-5 days in culture, the cells were harvested and counted (red bars). Error bars shown indicate the standard deviation. |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, cultured cells can be cryopreserved using the following procedure:
- Transfer detached cells to a conical tube and centrifuge
- Remove supernatant and resuspend in CryoDefend® Cell Lines Media
- Transfer cells to a cryovial and freeze at -80 °C overnight
- Transfer the frozen cryovial to liquid nitrogen
View Graphic Procedure
Reagents Provided
Reagents supplied in the CryoDefend® Cell Lines Media (Catalog # CCM019):
- 5 x 10 mL vials of CryoDefend® Cell Lines Media
Upon receipt, the CryoDefend® Cell Lines Media should be stored at = -20 °C in a manual defrost freezer. The media can be thawed at 2 °C to 8 °C or at room temperature. Thawed media can be aliquoted and stored at = -20 °C in a manual defrost freezer for up to 3 months. Thaw a fresh aliquot for each use. Avoid repeated freeze-thaw cycles.
Other Supplies Required
Reagents
- Cell culture media
Materials
- Cultured cells
- Cryovials
- 15 mL centrifuge tubes
- Serological pipettes
- Pipette and pipette tips
Equipment
- 37 °C and 5% CO2 incubator
- Centrifuge
Procedure Overview
R&D Systems Protocol for the Cryopreservation and Thawing of Stem Cells in CryoDefend® Cell Lines Media (Catalog # CCM019)
Cryopreservation of Cultured Cells
- Thaw CryoDefend® Cell Lines Media.
- Detach the cells from the cell culture dish.
- Transfer the cells to a 15 mL conical tube.
- Centrifuge at 200 x g for 5 minutes.
- Remove and discard the supernatant.
- Resuspendthe cells in CryoDefend® Cell Lines Media at 0.5-1.0 x 106 cells/mL.
- Transfer the cells to a cryovial.
- Freeze the cryovial at -80 °C overnight.
- Transfer the frozen cryovial to liquid nitrogen for storage.
Thawing of Cryopreserved Cells
- Warm appropriate cell culture media to 37 °C.
- Add pre-warmed culture media to the cryovial containing cryopreserved cells.
- Pipette up and down and as cells thaw, transfer the thawed cells to a 15 mL conical tube containing pre-warmed expansion media.
- Centrifuge at 200 x g for 5 minutes.
- Resuspend the cells in pre-warmed expansion media.
- Plate the stem cells at the desired density.
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