I'm a current medicine student and I'm in my first scientific initiation in my graduation. Currently, me and my professor are having some troubles with contamination in our cells. This is a bottle with HCT-116 cells (colorretal cancer). This cells were, previously frozen with contaminated fetal bovine serum. My professor said she thought she treated the cells, but this contamination keep returning. We supplement our media with penicilin and B- anfotericin at 1% each. The weird thing is: The bottle keeps healthy and, suddenly, from one day to another, these clouds appears and the bottle liquid looks cloudy. Could anyone, please, help me by advising what could I do or what fungus is this (If this is a fungus)?
At that magnification it is hard to see anything. Have you eliminated mycoplasma as a possible cause? Some bacteria and fungi are naturally resistant to some antibiotics. try using some with a different mode of action like tetracycline and nystatin or nipagen.
Hello Filipe Othmar
With correct working practice, antibiotics should not be used for the routine maintenance of cell lines. Unfortunately, HCT-116 cells were frozen with contaminated FBS, which could have been avoided. In the presence of antibiotics, contamination may be suppressed, but the phenotype or genotype of the cells could be altered. Relatively little is known about the effect of antibiotic use in cell culture. But I would like to mention that antibiotics are toxic and can alter the biochemistry of the cells. It can influence gene expression, and if you are to use these cells for carrying out genetic, genomic or other biological assays, then you are not going to obtain reliable results. Let me state that if an infection is not seen in culture because it has been suppressed but not eliminated using antibiotics, then all other cultures in the laboratory would be at risk.
So, my advice to you is to discard the culture because I feel the culture is not worth reviving. It is a hard decision to make, especially when you are dealing with a precious culture, but it would be wise to take such a decision.
I have attached an article below which you may find it useful.
Article Use antibiotics in cell culture with caution: Genome-wide id...
Best.
It is extremely difficult to completely eliminate bacteria from contaminated cells, and the contamination can return. Many labs have HCT116 cells. Therefore, if your cells are not invaluable (transfected for example), the best thing to do is to throw away your culture and request the cell line from a nearby lab.