The decision to use antibiotics to prevent contamination should be based on the individual researcher's needs and experience. The following table is a general guide for use of Gibco antibiotics in cell culture media. The concentrations given are for cell culture media containing serum; serum-free media generally require lower concentrations. Also available are solutions that use one or more antibiotics in conjunction with an antimycotic. For all media types, optimal concentrations of antibiotics and antimycotics should be determined empirically.
|Antibiotic||Recommended concentration||Antibiotic spectrum||Stability in culture media at 37°C|
|Anti-PPLO Agent Tylosin||10 – 100 μg/ml||Mycoplasma and gram positive bacteria||3 days|
|FUNGIZONE® (Amphotericin B)||0.25 – 2.5 μg/ml||Fungi and yeasts||3 days|
|Gentamicin Sulfate||5 – 50 μg/ml||Gram positive and gram negative bacteria and mycoplasma||5 days|
|Kanamycin Sulfate||100 μg/ml||Gram positive and gram negative bacteria and mycoplasma||5 days|
|Neomycin Sulfate||50 μg/ml||Gram positive and gram negative bacteria||5 days|
|Nystatin||100 U/ml||Fungi and yeasts||3 days|
|Penicillin G||50 – 100 U/ml||Gram positive bacteria||3 days|
|Polymyxin B Sulfate||100 U/ml||Gram negative bacteria||5 days|
|Streptomycin Sulfate||50 – 100 μg/ml||Gram positive and gram negative bacteria||3 days|
When an irreplaceable culture becomes contaminated, researchers may attempt to eliminate or control the contamination.
First, determine if the contamination is bacteria, fungus, mycoplasma, or yeast. Isolate the contaminated culture from other cell lines. Clean incubators and laminar flow hoods with a laboratory disinfectant, and check HEPA filters. Antibiotics and antimycotics at high concentrations can be toxic to some cell lines; therefore, perform a dose response test to determine the level at which an antibiotic or antimycotic becomes toxic. This is particularly important when using an antimycotic such as FUNGIZONE or an antibiotic such as tylosin, Cipro, or M-Plasmocin.
The following is a suggested procedure for determining toxicity levels and decontaminating cultures.
For Research Use Only. Not for use in diagnostic procedures.