Protocol for
Treating Mycoplasma-contaminated Cell Cultures with BM Cyclin
Remove
culture medium from culture vessels by aspiration. Add new culture medium
containing BM Cyclin 1 (4 μl of stock solution/ml, final concentration 10
μg/ml). Cultivate the cells for 3 days as usual.
Remove
culture medium, add new culture medium containing BM Cyclin 2 (4 μl of stock
solution/ml, final concentration 5 μg/ml). Cultivate the cells for 4 days,
repeat the above cycle twice. Cross-Contamination While not as common as
microbial contamination, extensive cross-contamination of many cell lines with
HeLa and other fast growing cell lines is a clearly-established problem with
serious consequences. Obtaining cell lines from reputable cell banks,
periodically checking the characteristics of the cell lines, and practicing
good aseptic technique are practices that will help you avoid
cross-contamination. DNA fingerprinting, karyotype analysis, and isotype
analysis can confirm the presence or absence of cross contamination in your
cell cultures. Using Antibiotics Antibiotics should not be used routinely in
cell culture, because their continuous use encourages the development of
antibiotic resistant strains and allows low-level contamination to persist,
which can develop into full-scale contamination once the antibiotic is removed
from media, and may hide mycoplasma infections and other cryptic contaminants.
Further, some antibiotics might cross react with the cells and interfere with
the cellular processes under investigation. Antibiotics should only be used as
a last resort and only for short term applications, and they should be removed
from the culture as soon as possible. If they are used in the long term,
antibiotic-free cultures should be maintained in parallel as a control for
cryptic infections.
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