Freezing Protocol

If freezing adherent cells

1. Remove culture medium. Wash the cell by DPBS (LB 001-02) or HBSS (LB 003-03, LB 003-04).
2. Cells, remove using Trypsin or Trypsin-EDTA for 1~3 minutes at 37°C. And completely remove Trypsin or Trypsin-EDTA by centrifugation cells at 80 xg for 2~3 minutes.
3. Resuspend cells gently in an appropriate volume of culture medium. And remove remaining Trypsin or Trypsin-EDTA by centrifugation cells at 80 xg for 2~3 minutes.
4. Resuspend cells gently in an appropriate volume of Cell Freezing Medium.
5. Dispense the cell suspension in 1~2 mL aliquots in plastic freezing vials or glass ampules.
6. Place vials in an insulated container and store in freezer for cooling rate -1°C/minute. Transfer vials to vapor phase of liquid nitrogen and store for 24 hours before transferring to liquid phase.
7. Cell viability count should be performed by melt the vial.

If freezing suspension cells

1. Remove culture medium by centrifugation cells at 80 xg for 2~3 minutes. Resuspend cells gently in an appropriate volume of Cell Freezing Medium.
2. Dispense the cell suspension in 1~2 mL aliquots in plastic freezing vials or glass ampules.
3. Place vials in an insulated container and store in freezer for cooling rate -1°C/minute. Transfer vials to vapor phase of liquid nitrogen and store for 24 hours before transferring to liquid phase.
4. Cell viability count should be performed by melt the vial.

Frozen cell`s culture

1. Remove vials from freezer and rapidly thaw in a 37°C water bath.
2. If DMSO or glycerol need remove, discard supernatant by centrifugation cells at 80 xg for 2~3 minutes. If DMSO or glycerol need not remove, resuspend cells of culture medium without centrifugation.
3. Transfer cells to a culture flask and slowly add the appropriate volume of growth medium.
4. Cultivate cell in incubator.

* In effort to cater to the individual needs of our clients, WelGENE provides custom products (classified as a “special order”) with variations made to the general specifications listed on the catalogue. Variations can be made in regards to 1. elimination of certain materials 2. alteration of chemical portions 3. addition of new materials and 4. change in pH.