Cell Freezing Medium – DMSO
Minimum Essential Medium Eagle
With 10% dimethyl sulfoxide (DMSO)
With 10% fetal bovine serum (FBS)

Catalog Number FM 001-01
Storage Temperature -5~-20°C

Product Description
Cell Freezing Media is used for freeze storing animal cells in liquid nitrogen. Cell Freezing Media contains serum supplements and freeze resistance regents such as dimethyl sulfoxide and glycerol. When freezing animal cells using the Cell Freezing Media, all cells should be trypsin treated or suspended in Cell Freezing Media after centrifuging prior to storage.

FM 001−01 contains 10% DMSO and 10% FBS in MEM base. Examine the culture for the absence of contamination, healthy growth confluency, etc.

Storage/Stability
The Cell Freezing Medium should be stored at -5~-20°C in the dark. Deterioration of the solution may be recognized by (1) precipitate or particulate matter throughout the solution, (2) cloudy appearance, (3) color change, and/or (4) pH change. Product label bears expiration date.

Biological Performance Characteristics
The growth-promoting capacities of Cell Freezing Medium are cell growth tested cell thawing after freezing. Growth rates are examined through three subculture generations and compared with parallel cultures grown in standardized control medium. Cells are counted and growth is plotted as a logarithmic function of time in culture, and seeding efficiency, doubling time, and final cell density are determined. During the testing period cultures are examined microscopically for a typical morphology and evidence of cytotoxicity.

Precautions
For In Vitro Use Only

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.