Preparation ratio of cryoprotectants for different cells (precautions for cryoprotectant preparation)
Cell cryopreservation solution is a special solution used to preserve cells in a low-temperature environment, with the main purpose of protecting cells from damage during freezing and thawing processes. Cell cryopreservation solutions typically contain both osmotic and non osmotic protectants, such as dimethyl sulfoxide (DMSO) and glycerol, which can penetrate into the interior of cells, reducing electrolyte concentrations inside and outside the cells, preventing water leakage from the cells, and thus reducing the formation of ice crystals.
Overview of Cell Cryopreservation Solution Configuration
The commonly used cell cryopreservation solution ratio is culture medium: serum: DMSO=7:2:1, which is suitable for most cell lines and can ensure a high survival rate of cells during cryopreservation. Cell lines that require long-term preservation or have special preciousness should be adjusted to a serum: DMSO ratio of 9:1, which can better protect the cells and ensure that they maintain good biological characteristics even after cryopreservation.
Proportion of different cell cryopreservation solutions
Most cells do not actually require 20% or 90% serum. They only need a complete culture medium containing 10% serum to prepare a solution containing 5% DMSO for cell cryopreservation, and some do not even require serum.
Some precautions for configuring frozen storage solution
✔ Choosing the appropriate cell state: Cells in logarithmic growth phase with a cell confluence degree of around 80% -90% should be selected for cryopreservation to ensure that the cells are in the optimal state for cryopreservation.
✔ Cell density: The cell density during cryopreservation should be appropriate, with a recommended density of 3 × 10 ^ 6 to 1 × 10 ^ 7 cells/mL. Low cell density may affect survival rate.
✔ Avoid prolonged exposure to room temperature: After adding the cell suspension to the freezing solution, the storage time at room temperature should be minimized as DMSO can cause significant damage to cells at room temperature.
✔ Use high-quality cryovials: Ensure the use of sterile and low-temperature resistant cryovials, and mark them properly during packaging for subsequent identification.
✔ The ratio of serum to DMSO: The commonly used ratio is 90% serum+10% DMSO or 55% basic medium+40% serum+5% DMSO, which can be adjusted according to specific cell types and experimental requirements.
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