Hi! We had a cell culture contamination for several weeks in different cell lines (HepG2, MCF-7, MDA). We have discarded all reagents, clean the waterbath and all the surfaces and started all over with other batch of cells, all seemed to be okay but 21 days later we have the same contaminant, we don't know what is.
The medium has str/pen but, in order to acelerate the posible contamination we starts using medium without str/pen and the result is the same, 15-20 days. They are moving and floating, and the medium color still red. They grow like a biofilm.
We have incubated the medium and FBS but nothing grew.
Add some pictures.
¿what could we do?
Hi Ana Maria
Persistent cell culture contamination can be frustrating. It sounds like you might have a mycoplasma or protozoan contamination, as they can form biofilms and are not inhibited by penicillin-streptomycin. To resolve this, consider:
Hope these steps help you resolve the issue. Good luck!
As the previous answer suggests, you should find out which is the contaminant you are dealing with. We have had mycoplasma contamination before, and we tried several reagents to try to eliminate it. In our hands the reagent from Roche worked in some of the cell lines; however, the time it took and the amount of work the treatments entailed favored our decision to purchase new cell lines for those that could not be treated.
Hi Ana Maria
You have mentioned that your media contains str/pen both of which are broad spectrum antibiotic. Try adding an antimycotic such as amphotericin B, to your medium before spending your time and resources for a possible mycoplasma contamination.
I also agree with Yogain Taank . selecting a specific antibiotic would work in your favor. Identify the antibiotic which might work for your assay, that might provide better and beneficial results to your lab settings!
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