SFM is used to cryopreserve all mammalian cells including stem cells and neuronal cells. It is for research use only. Not for use in animals, humans, or diagnostic procedures.
Cells cryopreserved using Serum-Free Freezing Medium show levels of viability and percent attachment (adherent cells) that are comparable to cells preserved in DMSO and FBS. Serum-Free Cell Freezing Medium can be used for both cells cultured in serum-supplemented growth medium as well as cells grown under serum-free conditions.
Store at -20°C. Once thawed, the product may be stored at 4°C for up to one month.
Cryopreservation may compromise cell quality and performance. Performance of the cells cannot be guaranteed after cryopreservation.
1. Harvest cells and spin them down.
2. Resuspend cells in cold SFM at a density of ~500,000 to 2,000,000 cells/ml. Work quickly.
3. Pipet 1 ml into each freezing vial and properly tight on the cap.
4. Place the cells (vials) in a Styrofoam® or propanol freezing canister and store them at -80ºC overnight. The cells should not be left at 80°C for more than 24 to 48 hours.
5. Transfer the cells to the vapor phase of liquid nitrogen (196°C) for longterm storage.
1. Warm the complete growth medium to 37°C prior to use with the cells.
2. Thaw a vial of cells cryopreserved in SerumFree Cell Freezing Medium by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water.
Thawing should be rapid (approximately 90 seconds).
3. Remove the vial from the water bath before the contents are completely thawed, and decontaminate by dipping in or spraying with 70% ethanol.
All of the operations from this point on should be carried out under strict aseptic conditions.
4. Dilute in warmed medium of choice at a ratio of 1 part sample in 10 parts medium.
5. Centrifuge the cell suspension at 1,000~2,000 rpm for 3~5 minutes at 4°C.
6. Aspirate the supernatant and resuspend the pellet in 2 mL of complete growth medium.
7. Count the cells; adjust the volume so that the cells are plated at the appropriate seeding density.