FM-1 (Cryoprotectant for Cell Culture)

Cells & Cell Culture/Cryopreservation

A cryoprotective solution containing serum- and protein-free medium components.
Suitable for cryopreservation of serum-free cultured cells.
This is a cryopreservation medium for serum-free cultured cells established by Ohno et al. in 1988. Cryopreservation of cells Generally, a medium containing dimethyl sulfoxide or glycerin and serum is widely used. As serum-free culture became more popular, media for cryopreservation in a serum-free state became desirable. Near It is known that the recovery rate of living cells is increased by using methyl cellulose as an alternative to serum. It came to be. This medium is based on this prescription. This cryopreservation medium can also be used to freeze cells cultured in normal serum. Usage is serum-free.
Storage method:
Dark place / moisture proof, 2-10 °C
Validity period: 1 year
Operation methods:
[Cell freezing method using FM-1]
Perform all operations aseptically.
This medium has twice the concentration of cryoprotectant.
(1) Add 5 mL of cell suspension suspended in the culture medium used for normal culture to 1 bottle (5 mL) of this medium.
Yes (1: 1). Alternatively, add this medium 1: 1 to the cell suspension.
(2) Fill (1) in a sterilized serum tube or ampoule, seal it, and put it in a styrofoam container for freezing.
Store at -80 ° C or below. Or freeze it in the program freezer.
Cell density is 106
Prepare to be above cells / mL.
Note) Methyl cellulose may precipitate in this medium, but its performance does not change.
[Method of thawing and reculturing frozen cells]
① Put the ampoule stored below -80 °C in hot water at 37 °C and shake it by hand to melt it rapidly.
To do.
(2) After thawing, open the package, dilute it 5 to 10 times with a normal culture solution, and settle for 3 to 5 minutes with a centrifugal force of about 1000 r.p.m.
vinegar.
(3) Discard the supernatant, add the required amount of normal culture solution, transfer to a culture vessel, and start culturing. Usually if necessary
Please wash the cells again with the culture solution of. After culturing overnight, if the culture solution is changed, the culture state will change.
References
1．Tadao Ohno, Cell Engineering，7(2)：171-172, （1988）
2．T. Ohno, K. Kurita, S. Abe, N. Eimori and Y. Ikawa：A simple freezing medium for serum-free cultured cells. Cytotechnology 1：257-260（1988）