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    5. Serum-free Cell Freezing Medium (SFM)

    Serum-free Cell Freezing Medium (SFM)

    Product Name
    Serum-free cell freezing medium (SFM)
    Cat.No.
    SF-CFM-01CL
    Product Use
    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.
    Application/Comments
    Cryopreservation
    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.
    Storage
    Store at -20°C. Once thawed, the product may be stored at 4°C for up to one month.
    FREEZING
    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.
    THAWING
    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.
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