Antibiotics and/or antimycotic agents are added to cell culture media as a prophylactic to prevent contamination, as a cure once contamination is found, to induce the expression of recombinant proteins, or to maintain selective pressure on trasfected DNA.
ATCC does not use antibiotics or antimycotics for routine cell culture. Long-term use of antibiotics or antimycotics may mask the presence of low levels of microbial or mycoplasma contamination. In addition, some antibiotics and antimycotics are toxic and may affect the recovery and proliferation of some cell lines.
However, one may elect to introduce antibiotics for short periods to primary cultures or as a safeguard while propagating specific valuable stocks to produce working stocks. If you do elect to use an antibiotic in your medium, ATCC recommends using a Penicillin-Streptomycin solution at a final concentration of 50-100 I.U./ml penicillin and 50-100 µg/ml streptomycin. ATCC offers a Penicillin-Streptomycin solution (ATCC® 30-2300 ™). This sterile solution can be added at 0.5 to 1 ml of solution per 100 mL of cell culture media for a final concentration of 50 to 100 I.U./mL penicillin and 50 to 100 µg/mL streptomycin.
While ATCC avoids use of the following two agents, commonly used concentrations are as follows. Gentamicin sulfate is an antibiotic and is used at 50 to 100 µg/ml culture medium. The antimycotic amphotericin B is used at 2.5 µg /ml culture medium. See Chapter 7 in Methods in Enzymology: Cell Culture, (1979) Vol. 58, W. B. Jacoby and I. H. Pasten, eds. (Academic Press, New York).
|Date Created||07/17/2012 07:10 PM
|Date Updated||01/23/2013 02:06 PM