Cryopreserved cells are fragile and require gentle handling. Thaw cells quickly and plate directly into complete growth medium. If cells are particularly sensitive to cryopreservation (DMSO), they are centrifuged to remove cryopreservative and then plated into growth medium.
The following are suggested procedures for thawing cryopreserved cells:
Direct Plating Method (This is what we use in our Lab)
- Remove cells from storage and keep on dry ice. Thaw quickly in a 37°C water bath.
- Plate cells directly with complete growth medium. Use 15ml of complete medium per 1 ml of frozen cells. Cell inoculums should be at least 3 x 105 cells/ml.
- Culture cells for 12 to 24 h. Replace medium with fresh complete growth medium to remove cryopreservative.