Thawing Cryopreserved Cells

What are the general Guidelines for Thawing Cryopreserved normal human Cells?


All cryovials should be stored in liquid nitrogen immediately upon arrival, unless thawed for cell culture.

We strongly recommend end user to follow all proper instructions before
working with the product. Refer to cell line specific instructions on the product page.


These are the general guidelines for thawing cryopreserved cells.

  1. Eliminate the cryovial in the liquid nitrogen storage tank.
  2. Thaw the cells immediately by putting the lower half of the vial into a 37°C water tub
    While agitating gently, remove after 60 minutes. Keep the cap out of the water to
    prevent contamination. There should still be a few ice crystals left after thawing (it is
    crucial not to over-thaw the cryovials as the presence of DMSO is toxic to the
    cells).
  3. Decontaminate the vial by spraying and wiping the exterior of the vial with 70%
    ethanol. From that point onwards, all operations should be rigorously carried out inside a
    biological safety cabinet in aseptic conditions.
  4. Gently re-suspend the cells at the vial and transfer the cell suspension to a 15 mL
    sterile conical tube containing 5 mL of pre-warmed, complete media using a sterile
    transfer pipette. 5. Centrifuge the cells in 200x gram for 3 minutes to pellet. Aspirate outside the supernatant
  5. Re-suspend the cell pellet in fresh, pre-warmed culture websites and transfer the cells
    Into a culture vessel. Gently rock the culture vessel to distribute the cells evenly. Table
    1 provides overall guidelines to the volume of culture media needed for a range of
    culture vessels.
    Note: The size of this culture vessel is exposed to the seeding density of the cell line (accessible
    On the corresponding cell line webpage”seeding density” section). Otherwise, we urge
    Thawing entire cryopreserved content to T25 flask with educated medium condition from
    The propagation segment of the item page.
  6. Incubate the culture at 37°C, 5% CO2, or some other advocated culture Atmosphere
    For the specific cell line. Incubate for 24 hours prior to processing the cells for downstream experiments.

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