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What should be considered to ensure efficient cell cryopreservation?
1) Slow freezing
Slow freezing allows cells to dehydrate gradually, preventing damage caused by the formation of large ice crystals. In contrast, the presence of large intracellular crystals can lead to damage and rupture of the cell membrane and cellular organelles.
2) Cell viability and concentration
Cells should be cryopreserved when they are in good growth condition, at a confluence of approximately 80–90%, and with viability exceeding 90%. Cells with poor viability exhibit very low post‑cryopreservation survival rates; therefore, it is essential to cryopreserve them during their peak proliferative phase.
3) Cryopreservation density
When the cell density during cryopreservation is less than 5 × 10^5 viable cells/mL, successful recovery is difficult.
4) Cryovial caps
The cap of the cryovial must be tightly sealed; otherwise, water may seep in during thawing in a water bath, leading to contamination.
5) Cryopreservation duration
Cells should not be stored long-term at −80°C.
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