Detailed steps for cell cryopreservation

Release time:

2023-03-23

Cell cryopreservation is the storage of cells at very low temperatures for thawing and recovery when needed. The following are the steps for general cell lyophilization:

Detailed steps for cell cryopreservation

Cell cryopreservation is the storage of cells at very low temperatures for thawing and recovery when needed. The following are the steps for general cell lyophilization:

1. Prepare lyophilization solution: prepare a cell lyophilization solution containing 10%-20%. Commonly used cell lyophilization solution components include glycerol, DMSO, FBS, etc. The specific formulation can be adjusted according to experimental needs.

2. Prepare cells: Culture cells to 80%-90% density, harvest cells, and wash 1-2 times with PBS or other buffers to remove cell debris and waste.

3. Prepare cell suspension: Add appropriate amount of lyophilization solution, suspend cells in cell suspension, and distribute them evenly into lyophilization tubes or other lyophilization containers.

4. Slow cooling: Place the cell lyophilization tubes or other containers and cool them slowly at -20°C isothermally, so that the cells gradually cool down to below -80°C.

5. Rapid freezing: Place the cell freezing tube or other container and freeze it rapidly in a container containing liquid nitrogen, so that the cells can be cooled down to below -196℃ very quickly.

6. Storage: Store the cell lyophilization tubes or other containers in liquid nitrogen until they are needed for thawing and recovery.

It is important to note that the procedure of cell lyophilization should be strictly followed to avoid any possible mishandling and contamination. And, when using cell cryopreservation, it is also necessary to pay attention to the steps and conditions of thawing. In general, the cell cryopreservation tubes or other containers should be placed in a 37°C water bath as soon as possible when thawing until the cells are completely thawed, and then they should be transplanted into a new culture dish with appropriate medium for culture.

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