Article Growth and Maintenance of Vero Cell Lines
Article Effect of Slow and Rapid Freezing Method on the Viability of...
I only find up to several hours at 4oC. You can google or other methods to search for what you need. Alternatively, you could freeze and check the viability at a series of time points at 4oC. Others may have a good idea.
Hello Akhil Anilkumar
A controlled gradual cooling method to -80 degree C at the conventional rate of 1°C/min must be followed by using the cryopreservation storage container like Mr. Frosty to avoid cell injury. Intracellular ice formation and osmotic injury are the main mechanisms leading to cell damage while freezing The gradual cooling at low temperatures may be explained by longer times required for the cells to pump out water in order to maintain osmolarity equilibrium with the extracellular medium when the temperature decreases. This is in line with the fact that extracellular osmolarity is determined by the temperature, and with the fact that the DMSO solution viscosity increases exponentially as the temperature decreases implying that the cooling rate should be slowed at lower temperatures to compensate for the lower water mobility.
Gradual cooling to -80 degree C dramatically increases the survival of cells in comparison to direct storage at -80 degree C from -4 degree C. Lowering the cooling rate to 0.5°C/min will result in an even higher survival rate of cells.
So, keeping the Vero cells in freezing media at -4 degree C for 15 hours and then moving to -80 degree C /liquid nitrogen will cause loss in cell viability when you thaw the cells the next time. You can do the viability check for these cells when you thaw them the next time.
Best.
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