StemXVivo Serum-Free MSC Freezing Media
R&D Systems, part of Bio-Techne | Catalog # CCM016
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, MSCs can be cryopreserved for long-term storage using the following protocol:
- MSCs are detached from the culture dish
- MSCs are resuspended in StemXVivo® Serum-Free MSC Freezing Media
- Frozen, cryopreserved MSCs are stored in liquid nitrogen
Reagents supplied in the Human StemXVivo® Serum-Free Mesenchymal Stem Cell Freezing Media (Catalog # CCM016):
- 50 mL of serum-free MSC freezing media
Reagents
- Human StemXVivo® Serum-Free MSC Expansion Media (Catalog # CCM014 or equivalent)
- TrypLE™ Express (Invitrogen® or equivalent)
Materials
- Human MSCs
- Tissue culture flasks
- 15 mL centrifuge tubes
- Cryogenic storage vials
- Pipettes and pipette tips
- Serological pipettes
Equipment
- 37 °C and 5% CO2 incubator
- -80 °C freezer
- Liquid nitrogen storage
- Centrifuge
- Hemocytometer
- Inverted microscope
- 37 °C water bath
This protocol has been tested using bone marrow- and/or adipose tissue-derived MSCs. If using a different tissue source or cell line, the protocol below may need to be modified.
Freezing
Detach the MSCs from the culture dish using a dissociation solution.
Transfer the dissociated cells to a 15 mL conical tube.
Centrifuge the MSCs at 200 x g for 5 minutes.
Resuspend the cell pellet in cold StemXVivo® Serum-Free MSC Freezing Media.
Transfer the MSC suspension to a cryovial immediately.
Place the cryovial in a cryogenic container at -80 °C overnight.
Store the frozen cryovial in liquid nitrogen.
Thawing of Cryopreserved Stem Cells
Remove the frozen vials from the liquid nitrogen, and rapidly thaw in a 37 °C water bath.
Transfer the cells into a 15 mL centrifuge tube containing pre-warmed StemXVivo® Mesenchymal Stem Cell Expansion Media.
Centrifuge the MSCs at 200 x g for 5 minutes.
Resuspend the cells in serum-free complete growth medium and plate the MSCs in desired culture conditions.
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